Дисертації з теми "Legal status of stem cells"

Les cellules souches en tant que produits cellulaires à finalité thérapeutique (PCT) ou en tant que médicaments de thérapie innovante (MTI) dans le cadre de la médecine régénératrice ont révolutionné la médecine du XXIe siècle. Face aux découvertes récentes de nouvelles cellules souches créées par les chercheurs (parthénotes, cellules souches clonées, cellules iPS), d’autres possibilités de thérapie régénérative surgissent au fil du temps.Le droit, qui a toujours accompagné l’évolution scientifique et technique de la thérapie cellulaire depuis le XVIIe siècle, doit être plus que jamais présent pour protéger l’être humain qui se prête aux nouveaux traitements ou à l’expérimentation. L’évolution historique de cette révolution thérapeutique nous permet de montrer l’importance de la réflexion juridique et éthique pour le progrès scientifique. Des questionnements anciens, comme le statut de l’être prénatal et l’autorisation de cryopréservation des tissus ou des cellules autologues, ressurgissent face à la présence de cellules souches humaines embryonnaires surnuméraires et aux succès de la thérapie régénérative. Des traitements tératogènes et des épisodes de maltraitance des femmes en cours de grossesse ont détruit ou endommagé des milliers d’enfants à naître. Une reconnaissance de la vie prénatale est proposée dans certaines circonstances pour protéger l’embryon et le fœtus avant leur naissance
Stem cells as cellular products for therapeutic purposes (PCT) or as advanced therapy drugs (ITNs) within the framework of regenerative medicine have revolutionized the medicine of the 21st century. Faced with recent discoveries of new stem cells created by researchers (parthenotes, cloned stem cells, iPS cells), other possibilities for regenerative therapy are emerging over time.The law, which has always accompanied the scientific and technical development of cell therapy since the 17th century, must be more present than ever to protect human beings who lend themselves to new treatments or to experimentation. The historical development of this therapeutic revolution allows us to show the importance of legal and ethical reflection for scientific progress.Old questions, such as the status of the prenatal being and the authorization for cryopreservation of autologous tissues or cells, are re-emerging in the face of the presence of supernumerary human embryonic stem cells and the success of regenerative therapy. Teratogenic treatments and episodes of child abuse during pregnancy have destroyed or damaged thousands of unborn children. Recognition of prenatal life is offered in certain circumstances to protect the embryo and fetus before birth

Human embryonic stem cell research would seem to offer the prospect of developing a greater understanding of, and potential therapies for, common degenerative diseases such as diabetes mellitus, Alzheimer’s and Parkinson’s disease. Despite the fact that some Irish institutions engage in such research, Ireland is one of the few countries in Europe which has failed to produce any relevant regulatory framework or legislation. This is largely because embryo research and its regulation remain mired in conflicting socio-political values and interests, despite the fact that the in vitro human embryo is not afforded any legal protection under the Irish Constitution. This thesis seeks to examine the current Irish legal lacuna in relation to embryos and embryonic stem cell research. The first of the three papers making up the core of this thesis reviews the background to the moral, legal and social factors that have contributed to the extant Irish position. A description of the divergent policies enacted in other jurisdictions is also given to outline possible policy options which may be considered by Ireland in the future. The views of relevant stakeholders on the impact of the regulatory lacuna are explored in the second paper through a series of semi-structured interviews. These interviews highlight a surprising level of consensus on the need for the Irish legislature to act and introduce regulations to provide certainty, in one way or the other, in this area of scientific innovation. A procedural mechanism is proposed in the third paper which could allow the development of policy and concomitant regulation in Ireland in this area. It is hoped that the procedural process and resultant framework would be sufficiently inclusive as to be acceptable to the majority of people in Irish society. In conclusion, it is argued that it is undesirable that a modern pluralist democracy (as Ireland aspires to be) should regard legislative inertia and non-regulation as the preferred method of dealing with morally challenging scientific endeavour. Instead, appropriate procedural mechanism should be utilised to allow for the development of apposite policies.

In adults blood stem cells, called haematopoietic stem cells (HSC), give rise to all blood cells throughout life. The origin and biology of HSCs during embryo development has been an intensely studied topic. Definitive HSCs are generated intra-embryonically in the aorta-gonad-mesonephros (AGM) region of the mid-gestation embryo. Recent research revealed that HSCs emerge through multistep maturation of precursors: proHSC → preHSC I → preHSC II → definitive HSC (dHSC). A hallmark of the HSC emergence is the appearance of intra-aortic haematopoietic clusters that are considered to be sites of haematopoiesis. It was shown in vitro that the E11.5 HSCs are slowly cycling compared to progenitor cells. However, cell cycle status and its role during early HSC development remain unclear. Here I used Fucci transgenic mice that enable in vivo visualisation of the cell cycle. Functional and phenotypic analysis showed that in the early embryo the proHSC precursors cycle slowly, whereas committed progenitors are actively cycling. Meanwhile the preHSC I precursors arising in the E10.5 AGM region become more rapidly cycling. They are located closer to the luminal cavity of the dorsal aorta, while their ancestors, the proHSCs, are slowly cycling and are located at base of the clusters. Furthermore, in the mid-gestation embryo the preHSC I become slowly cycling and are closer to the endothelial lining of the aorta, while they give rise to the actively cycling preHSC II that are located to the luminal area of the artery. Finally, definitive HSCs are mainly slowly cycling at this stage like their foetal liver counterparts. As expected, HSCs in adult bone marrow are mainly dormant. The data suggest that transition from one precursor type to another is accompanied by distinct changes in cell cycle profile and that HSCs become progressively quiescent during development. To test the role of cell cycle in HSC maturation, we used inhibitors against signalling pathways known to play important roles in HSC development. Notch inhibitor affected the cell cycle status of haematopoietic precursors, by possibly promoting them to rapidly proliferate and potentially blocking the maturation from preHSC I to preHSC II precursors. Shh antagonist had the opposite effect and enhanced the HSC activity from the preHSC I precursors. Altogether these results suggest that the cell cycle status plays an important role in the HSC development. A better understanding of the molecules that control this process will allow us to optimize the culture condition for generation of functional HSCs in the laboratory.

Thesis (DPhil (Philosophy))--University of Stellenbosch, 2007.
Should surplus embryos which are destined to be discarded be protected at all cost, to the extent that they cannot contribute to medical knowledge - knowledge which could benefit society at large? Are embryos people or merely items of property? Different moral theories address these questions in different ways. Deontologists argue that the end never justifies the means and that the right not to be killed is more fundamental than the obligation to save. Utilitarians, on the other hand, argue that certain criteria should be met before moral significance can be contributed to an entity. The question of the moral status of the embryo is, as my discussion will show, one of the most widely discussed issues in the history of bioethics. Extensive literature exists on the topic. This study holds that an Ethics of Responsibility (ER) should by applied when answering the questions posed above as it encourages one to accept responsibility for the choices or decisions made and to defend them accordingly. I have endeavoured to answer the question of the personhood and rights of the embryo within the framework of the Ethics of Responsibility. Although these concepts overlap in many ways they remain central to the debate surrounding the sanctioning or prevention of the use of human embryonic stem cells in research. After identifying the micro-issues surrounding the human embryonic stem cell debate and explaining why both the deontologist and utilitarians fail to provide any adequate answers in this respect, I turn my attention to macro-issues such as safety concerns surrounding the usages and storage of stem cells. Commercialization, power issues, accessibility and the allocation of limited resources are also examined. Living in a society such as South Africa one cannot be blind to the inequalities of our health system. On a macro level I cannot but conclude that stem cell research does not seem to be a viable exercise within the South African context. South Africa faces a health care crisis far greater than the benefits stem cell research currently has to offer. However, the need still exists for a policy to guide future lawmakers who might need to address stem cell research and to guide decisions and actions. This brings me to my final chapter, namely proposing a morally justified policy for South Africa. I propose a policy which respects and values the autonomy of the progenitors’ choices (provided they have not been coerced) and which focuses on the beneficence of the greater society. Furthermore, it is paramount that the goal of any stem cell research should be for therapeutic use ONLY. Before commencing with the extraction of the stem cells, scientists should be obligated first to present convincing evidence that they have tried alternative ways to reach the same result. Once this has been proven, a regulatory body could issue the scientist/team with a license to undertake the specific research with a specific therapy as goal in order to prevent abuse. If they are found guilty of any unethical conduct their licenses should be revoked and an investigation launched.

Rapid progress of AI technology in the life science area is observed in recent years. Convolutionalneural network (CNN) models were successfully applied for the localization and classification of cellson microscopic images. Induced pluripotent stem cells are one of the most important innovations inbiomedical research and are widely used, e.g. in regenerative medicine, drug screening, and diseasemodeling. However, assessment of cell cultures’ quality requires trained personnel, is timeconsumingand hence expensive. Fluorescence microscope images of human induced pluripotentstem‐hepatocytes (hiPS‐HEPs) derived from three human induced pluripotent stem cell (hiPSC) lineswere taken daily from day 1 until day 22 of differentiation. The cells from day 1 to 14 were classifiedas ´Early differentiation´, and above day 16 as ´Late differentiation´. In this study, it wasdemonstrated that a CNN‐based model can be trained with simple fluorescence microscope imagesof human induced pluripotent stem‐hepatocytes, and then used to predict with high accuracy(96.4%) the differentiation stage of an independent new set of images.

The rapidly evolving biosciences increasingly rely on the analysis, manipulation and reproduction of the human body. In the health setting, novel biotechnologies offer new methods/avenues for the investigation of wellbeing and the treatment of illness, but they do not just expand the clinician’s toolbox, they increase the very scope of her work. By offering new (and formerly invisible) measures for health, they have created new categories of illhealth (ie: expanding the ways in which humans can be classified as abnormal, unhealthy, or diseased). In doing so, they contain huge marginalising potential. And they are evolving at a pace that the law cannot match. Given this, important questions arise such as: What institutions are acting in this field and what is guiding them? How is health-related research being encouraged and regulated? How does the human subject figure in the bioeconomy? What values are we claiming and vindicating under existing regulatory regimes? What values ought we be emphasising bearing in mind social needs and individual rights? The body of work that forms this submission represents five years of socio-legal research and evolving thought on the topic of how values inform the law and are operationalised through the law and legal institutions. While the publications relied on are diverse, they all pursue small facets of this value inquiry. The first theme addressed – international values and actors – is composed of three papers which explore broad internationally shared values claimed in legal instruments such as the Universal Declaration on the Human Genome and Human Rights and the Universal Declaration on Bioethics and Human Rights, and institutions such as UNESCO and the EPO. A range of values emerge from these. Papers under the second theme – human participation in health research – explore how we access and use the human body in the modern biosociety/bioeconomy, and how we might better encourage subject participation in, and equitable benefit from, the biomedical research setting. Focusing on population biobanking, it assesses who has rights in the body and what those rights are, and how the existing environment interacts with our claimed values. Papers under the third theme – encouraging stem cell research in Argentina – explore governance instruments and their significance for realising claimed or desired values. These papers are informed by original empirical work conducted in Argentina over a 24-month period during which the Argentine government grappled with the realities of the new biosociety and the (perceived) need to facilitate bioscience research and medical treatment using human tissue. While these papers represent only part of the scholarship deriving from this project, they deploy new evidence on the existing environment and the way forward in that jurisdiction. As argued in the Critical Review, these publications form a broadly coherent and farranging body of interdisciplinary work which persistently questions the link between law and values and how we govern modern bioscience. While there are necessarily descriptive elements, the whole is critically analytical and normatively suggestive. In addition to summarising the aims, objectives, methodology, results and conclusions of these works, and indicating how they form a coherent body of work, the Critical Review goes further. Drawing on evolving thinking and recent scholarship, it argues for a regime less reliant on instruments and more reliant on expert institutions informed by, and charged with protecting, socio-moral values informed by the human rights paradigm.

Osteoblasts (OST) provide strong intrinsic growth modulatory activities on hematopoietic stem and progenitor cells via different mechanisms that include secretion of growth factors, and cellular interaction. Previously we showed that medium conditioned by mesenchymal stromal cell (MSC)-derived osteoblasts (M-OST) improve the expansion of cord blood (CB) CD34+ cells. I hypothesize that the hematopoietic supporting activity of M-OST would vary as a function of their maturation. This was tested by producing osteoblast conditioned media (OCM) from M-OST at distinct stages of maturation, and testing their growth regulatory activities in CB CD34+ cell cultures. My results showed that some of the growth promoting activity of OCM on CB cells are not dependent on the maturation status, while others are and those are largely independent of Notch signalling. In conclusion, these results provide further evidence that osteoblasts release factors that can promote the growth of immature CB progenitors in a Notch-independent way.

Within many communities and religions, including the LDS community, there is some controversy surrounding the use of stem cells – particularly embryonic stem cells (ESC). Much of this controversy arises from confusion and misconceptions about what stem cells actually are, where they come from , and when life begins. The theology of the Church of Jesus Christ of Latter-day Saints has interesting implications for the last of these considerations, and it becomes less a question of “when does life begin” and more an exploration of “when does personhood begin” or “when does the spirit enter the body.” With no official Church stance, statements from Church leaders vary on this topic, and this first section of the thesis explores the philosophical and practical meaning of personhood with a biological background intended for those not familiar with the origin or uses of stem cells.The second portion of the thesis explores possible cloning technologies. Recent events and advances address the possibility of cloning endangered and extinct species. The ethics of these types of cloning have considerations uniquely different from the type of cloning commonly practiced. Cloning of cheetahs (and other endangered or vulnerable species) may be ethically appropriate, given certain constraints. However, the ethics of cloning extinct species varies; for example, cloning mammoths and Neanderthals is more ethically problematic than conservation cloning, and requires more attention. Cloning Neanderthals in particular is likely unethical and such a project should not be undertaken. It is important to discuss and plan for the constraints necessary to mitigate the harms of conservation and extinct cloning, and it is imperative that scientific and public discourse enlighten and guide actions in the sphere of cloning.

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By establishing a foundation for final decisions of merit, judicial authority uses several elements, such as language, interpretation and discourse, which will comprise the content of the sentence or judgment, culminating in the grounds or lack of grounds for the request. However, in collegiate judgments, even if the conclusion of decisions pronounced by judges is the same, the votes rendered take different paths, characterized by the oneness of foundations chosen, even in cases of decisions rendered unanimously. Although there is no uniformity of understanding, the conclusions presented are considered acceptable, with the interpretation made by winning votes prevailing. This dissertation intends to demonstrate the presence of extrajudicial elements in the formation of the judge's conviction, characterized and raised by subjectivity. The certainty of objectivity functions as a cloak spread over subjectivity. The demonstrative method and study of arguments will be used according to the structure elaborated by Chaïm Perelman and also from the perspective of legal and non-legal argumentation found in the judgment of ADIn 3510/DF on the use of embryonic stem cells obtained from human embryos produced by in vitro fertilization and not used in the respective procedure. The study will consist of showing there is a merger of objective and subjective elements in the foundation related to the belief of the legal exegete. It shall be concluded that subjectivity is tied to experience and culture found in the foundation for the votes, encompassing valued judgments and their arbitrary (discretionary) application, implicitly containing intuition. The scope of the dissertation will be a case study (ADIn 3.510/DF) and the validity of the procedure, starting with the choice of a sample of the vote population emanating from the Ministers of the Federal Supreme Court, and checking whether it constitutes an outline of the population of decisions handed down by jurisdictional entities
O julgador, ao fundamentar as decisões terminativas de mérito, utiliza-se de vários elementos tais como a linguagem, a interpretação e o discurso, os quais comporão o conteúdo da sentença ou acórdão, culminando na procedência ou improcedência do pedido. Todavia, nos julgamentos colegiados, ainda que a conclusão das decisões proferidas pelos julgadores seja a mesma, os votos exarados trilham caminhos diversos, caracterizando-se pela não unicidade dos fundamentos escolhidos, mesmo nos casos de decisões prolatadas por unanimidade. Embora ausente a uniformidade de entendimento, as conclusões apresentadas são consideradas aceitáveis, prevalecendo a interpretação feita pelos votos vencedores. Esta dissertação pretende demonstrar a presença de elementos extrajurídicos na formação da convicção do julgador, caracterizados e deflagrados pela sua subjetividade. A certeza da objetividade atua como um manto estendido sobre a subjetividade. Será utilizado o método demonstrativo e o estudo dos argumentos de acordo com a estrutura elaborada por Chaïm Perelman e também sob a ótica da argumentação jurídica e não-jurídica constantes do julgamento da ADIn 3.510/DF, sobre a utilização de células-tronco embrionárias obtidas de embriões humanos produzidos por fertilização in vitro e não utilizados no respectivo procedimento. A investigação consistirá em evidenciar que há fusão de elementos objetivos e subjetivos na fundamentação, relacionados à crença do exegeta jurídico. Concluir-se-á que a subjetividade está ligada à experiência e cultura, contidas na fundamentação dos votos, englobando juízos valorativos e sua aplicação arbitrária (discricionária), contendo implicitamente a intuição. A dissertação terá como escopo um estudo de caso (ADIn 3.510/DF) e a validade do procedimento, partindo-se da escolha de uma amostra da população de votos emanados pelos Ministros do Supremo Tribunal Federal, verificando-se se constitui um traço da população de decisões proferidas pelos órgãos jurisdicionais

La découverte des cellules souches embryonnaires et de leur immense potentiel thérapeutique a fait naître de grands espoirs. De nouvelles thérapies révolutionnaires pour traiter certaines des maladies les plus graves dont souffre l'humanité sont désormais envisageables. Le traitement de la vie à son stade le plus précoce est mis en cause. Le statut juridique reconnu au foetus et à l'embryon humain a des répercussions directes sur le domaine de la recherche et sur leur utilisation à des fins thérapeutiques. Nous avons examiné l'état du droit canadien quant au statut juridique du foetus et de l'embryon. De cette étude, nous avons constaté l'incertitude qui prévaut au Canada quant à leur statut. Par la suite, nous avons étudié les différentes normes canadiennes établies pour encadrer l'utilisation des cellules souches embryonnaires à des fins thérapeutiques et nous les avons analysées et comparées pour faire ressortir leurs similitudes et leurs différences. II est ressorti de notre analyse que les textes canadiens se rejoignent généralement sur l'essentiel et qu'il y a eu peu de changements de 1993 à aujourd'hui, en regard des activités de recherche interdites au Canada. Puis, nous avons examiné les systèmes normatifs applicables à ces recherches à l'étranger, soit aux États-Unis et en GrandeBretagne. Nous avons effectué une analyse comparative des trois systèmes normatifs étudiés, en évaluant différents paramètres communs à ces systèmes. Il est ressorti de cette analyse, que la Grande-Bretagne est le pays le plus libéral relativement à ces domaines de recherche, que les États-Unis sont les plus conservateurs sur ces questions et que le Canada se situe entre les deux.
The embryonic stem cells discovery and the immense therapeutic potential glven to them has created big hopes in the world of today. The appearance of new revolutionary therapies to treat sorne of the most serious known diseases are now conceivable. However, the treatment of life to its earliest stage is questionned. The legal status recognized to the foetus and the embryo has, in fact, a direct effect to the research area and industry as weil as to its therapeutic use. Therefore, we have examined and studied the CUITent canadian law with respect to the legal status of the foetus and embryo. Following this study, we have noticed the uncertainty that prevails in Canada concerning the said legal status. Afierwards, we have examined ail the different canadian norms and regulations already established regarding the use of embryonic stem cells for therapeutic ends. We also did the comparaison between those norms and regulations so as to see their differences and similarities. It appears from our analysis that ail the canadian litterature generally treat the subject in the same way and that there have been few changes from 1993 up until now with respect to the forbidden researchs activities in Canada. We also have analysed the foreign law standards and regulations in United States and Great Britain concerning those forbidden researchs activities. We did the exercise of comparing the state of the law in these three countries with different parameters. It emerges from that that Great Britain is the most liberal country, United States being the most conservative and Canada being in between them.
"Mémoire présenté à la Faculté des études supérieures en vue de l'obtention du grade de Maîtrise en droit (L.L.M.) Option recherche"

碩士
國立交通大學
科技法律研究所
94
A stem cell is a cell from the embryo, fetus, or adult that has a unique capacity to renew itself and to give rise to specialized cell types. It also can differentiate into any type of tissue or organ, presenting the enticing prospect that they could one day be used to replace diseased or damaged cells and tissue. In 1998 cultures of human pluripotent stem cells were first created in the laboratory of James Thomson at the University of Wisconsin-Madison (Science 282, 1145; 1998). Research using stem cells is an extremely active area of current biomedical inquiry. Research efforts have focused on spinal cord injury, multiple sclerosis, Parkinson’s disease, Alzheimer’s disease, diabetes, and other diseases or conditions. Scientists hope to use specialized cells to replace dysfunctional cells in the brain, spinal cord, pancreas, and other organs. Other potential applications for human stem cell cultures include uses for studying fundamental processes of human development or for toxicological testing and drug design. Non-human animal stem cell lines may also be used to produce genetically modified animals. However, the rapid march of stem cell research doesn’t give society much time to figure out a coherent response to its discoveries, both because of its extraordinary promise and because of relevant legal and ethical issues. Controversy surrounds the derivation of stem cells from human embryos and fetuses. In order to derive or extract the stem cells found within the embryo, the embryo is destroyed in the removal process. Given the moral implications of this extraction and the sanctity attached to embryos by many groups and individuals around the world, it is unsurprising that the question of how the research should proceed has spawned an energetic debate. This study employs the comparative method based on Laws and Regulations in Taiwan, foreign literature review and Anglo-American Law to discuss the likely disputes on using and regulating embryonic stem cells. Furthermore, there are many unresolved ethical issues related to the clinical and experimental use of adult stem cells such as umbilical cord blood. These issues include determination of ethical procedures for informed consent and institution review border for adult stem cell donation to public banks, to private banks, and for research and treatment. For example, legal and ethical issues related to privacy, confidentiality, and ownership of cord blood units are complex and controversial. There is also considerable debate regarding the ethics of commercial cord blood banking, particularly related to the availability of this potentially valuable resource for clinical use and research. Finally, the legal systems concerning patentability of stem cells are different in the U.S., Europe and Taiwan. This study discusses the advantages and disadvantages of grant of patent for stem cells, the feasibility and limitation to the protection for the patent of stem cells.

碩士
國立高雄第一科技大學
科技法律研究所
97
Technology advancement and medical devices technology development have brought great contributions to human civilization. Stem Cell, extracted from the Umbilical Cord Blood, is one of the significant discoveries in our history without doubt. Cord Blood was once deemed medical waste and of no value at all, while it is nowadays medical treasure due to its stem cells. Therefore, his study will first clarify the ownership of the Cord Blood for further discussion about its related legal issues. In recent years, Cord Blood Registry (Cord Blood Banks) keep advising and promoting the benefits of preserving the Cord Blood. Many parents-to-be are wondering whether to preserve the Cord Blood for future utilization. If they determine to preserve the Cord Blood, they would next have to conduct a Contract with the Cord Blood Bank, public or private. What exactly is the nature of the Contract? How do the parties arrange the articles in the Contract? Since our Department of Health, Executive Yuan (DOH) has drafted the “Template of Standardized Contract for the Cord Blood Preservation”, this study will present several critical discussions to examine the articles of the DOH Template. This study will also provide concrete suggestions for parents-to-be to refer to when they conduct a Contract of Cord Blood Preservation in the future. This study would in addition introduce the regulation models of Cord Blood Banks in European Union and the United States, and review the current management regulation of Cord Blood Banks in Taiwan. This study will then provide for the Government’s reference regarding laws and regulations applicable to practical management of Cord Blood Banks.

碩士
國立交通大學
生物科技學系
101
Mesenchymal stem cells (MSCs) are the multipotent stem cells that can give rise to mesenchyme-lineage cells, such as osteoblasts, adipocytes, chondrocytes and myoblasts. MSCs can be isolated from various tissues including bone marrow, adipose tissue, and umbilical cord blood etc. Recently, MSCs have been represented as a promising cell source for further cellular therapy. In spite of the strong self-renewal property, MSCs undergo cellular senescence during expanded cultivation in vitro. It is well-known that the oxidative stress plays an important role for cell aging. In this study, we focused on the relationship between cellular oxidative status and cultivated senescence in bone marrow-derived MSCs (BMMSCs) and cord blood-derived MSCs (CBMSCs). We examined the cellular oxidative stress by culturing cells with antioxidants, kinetin and epigallocatechin-3-gallate (EGCG). To understand the effects of antioxidants supplementation on MSCs, the results of growth status, reactive oxygen species (ROS) value, and lipid peroxidation were analyzed in BMMSCs and CBMSCs. We found that kinetin and EGCG treated MSCs expressed similar surface marker profiles with normal culture condition. The results also showed that the lower oxidative status, the higher catalase activity, and the longer lifespan could be found in kinetin cultivation than control condition in both types of MSCs. However, there were no obvious differences between MSCs cultures with or without EGCG addition. In differentiation test, the osteogenesis of CBMSCs could be enhanced by kinetin and the adipogenesis could be inhibited by EGCG in both types of MSCs. Moreover, kinetin but not EGCG decreased the anti-aging marker β- galactosidase in late passage of BMMSCs and CBMSCs. These results indicated that kinetin could be a useful antioxidant to reduce cell senescence during MSCs expansion.

碩士
東吳大學
法律學系
93
Medical research and clinical practice throughout the latter half of the twentieth century brought human beings to the brink of curing many devastating diseases, especially the malignancies, that have plagued, the most promising one of which would be the umbilical cord stem cell related biotechnology. The umbilical cord blood stem cell gains prevailing advantages over bone marrow and fetal stem cell in both technical and ethical aspects, which makes the business community recognize that umbilical cord blood cell transplant technology could yield significant financial profit. These profit-making enterprises have established autologous umbilical cord blood storage services, inviting parents to bank their umbilical cord blood for possible future use all over the developed world. In Taiwan it has stimulated a ready market for blood storage, and several companies offering private storage and some charity community offering public umbilical cord blood banks now exist, which make the prevalence of umbilical cord blood stem cell storage rate estimated up to 4 per 100 births in this country. The umbilical cord stem cells, used to be regarded as delivery wastes, appear in itself legally a brand new issue both on personal rights and property rights which have never been well elaborated in the past. Disputes are certainly to arise since the umbilical cord blood stem cell so valuable a kind of substance, which urges us to pay more attention. This article elaborates the biological characteristics, clinical applications, and future scientific implications of stem cell, with special emphasis on the unique properties of umbilical cord blood stem cell, in the leading Chapter 2, which will help to make better understanding on differences between the umbilical cord blood stem cell and other stem cell origins. The status quo and the high prevalence on umbilical cord blood banking services, private or public, in Taiwan is briefly expounded in Chapter 3. The general concepts regarding personal rights of excised human organ and tissue are reviewed concisely in Chapter 4, and a novel canvass “personal nature” is introduced wherein to appropriately characterize an interim category transitionary from the property rights to personal rights. This article delimits precisely the demarcation line of “personal nature” and “personal rights” by examining the proposed formal and essential requirements. The ownership of the umbilical cord blood stem cell and its derivatives is discussed in Chapter 5, where the wrong classic concepts are fully presented. Meanwhile this article address that mothers have the entitlement to ownership of the umbilical cord blood stem cell by all the legal, anatomical, biological, and genetic aspects. Also this article holds that the derivative inducing process of umbilical cord blood stem cell is not kind of natural breeding as coded in Article 69 and 766 of Civil Code, but that of the processing work as coded in Article 784 of Civil Code. Chapter 6 of this article examined the validity of the clauses of umbilical cord blood deposit contracts. Lack of proper regulations is described in Chapter 7. Finally this article makes some proposition and advice for the umbilical blood banking community

碩士
國立中央大學
哲學研究所碩士在職專班
97
The important ethical issues of the life right to life and morals status of the embryo are trickered by the fast progress of human embryonic stem cells research. These ethical disputes must be analyzed in greater details in order to make a proper judgment of the practice of embryonic stem cell research. Therefore, this thesis first gives an analysis of the terms and concepts involved in the debate. It presents Peter Singer’s argument against the thesis of sanctity of life and Mary Anne Warren’s criteria of personhood. Both are single criterion for moral status and criticized by the latter Warren as inadequate to cover the diverse field of moral status. Warren proposes a multi-criteria account of moral status, but offers insufficient status to embryos in accordance with our moral experience. A Confucian proposal is put forward to mitigate some of its pitfalls. According to Confucianism we are endowed with a natural concern of the sufferings of others. Thus, a person is a moral agent and must act morally according to her feeling towards the suffering of others. The moral status of embryo can thus be established by the moral expression of humanity from the moral agency so that it becomes sufficient and reasonable to place proper moral judgment in embryonic stem cells research. We express the hope that laws and regulations could be adjusted in time while new technology is innovated in embryonic stem cells research. At the same time, we also seek alterative methods for the development of stem cell lines to avoid the destruction of embryos in the process. Then the research of embryonic stem cells could promote the value and quality of human life.

RESUMO: Actualmente, a única possibilidade de cura para doentes com adenocarcinoma do pâncreas (PDAC) é a ressecção cirúrgica, no início deste estudo, perguntamo-nos se os predictores clínico-patológicos clássicos de prognostico poderiam ser validados em uma grande cohort de doentes com cancro do pâncreas ressecável e se outros predictores clínicos poderiam ter um papel na decisão de que doentes beneficiariam de ressecção cirúrgica. No capítulo 2, observamos que até 30% dos doentes morrem no primeiro ano após a ressecção cirúrgica, pelo que o nosso objectivo foi determinar factores pré-operatórios que se correlacionam com mortalidade precoce após ressecação cirúrgica com recurso a um instrumento estatisticamente validado, o Charlson-Age Comorbidity Index (CACI), determinamos que um CACI score superior a 4 foi preditivo de internamentos prolongados (p <0,001), complicações pós-operatórias (p = 0,042), e mortalidade em 1 ano pós- ressecção cirúrgica (p <0,001). Um CACI superior a 6 triplicou a mortalidade no primeiro ano pós-cirurgia e estes doentes têm menos de 50% de probabilidade de estarem vivos um ano após a cirurgia. No capítulo 3, o nosso objectivo foi identificar uma proteína de superfície que se correlacionasse estatisticamente com o prognostico de doentes com adenocarcinoma do pâncreas e permitisse a distinção de subgrupos de doentes de acordo com as suas diferenças moleculares, perguntamo-nos ainda se essa proteína poderia ser um marcador de células-estaminais. No nosso trabalho anterior observamos que as células tumorais na circulação sanguínea apresentavam genes com características bifenotípica epitelial e mesenquimal, enriquecimento para genes de células estaminais (ALDH1A1 / ALDH1A2 e KLF4), e uma super-expressão de genes da matriz extracelular (colagénios, SPARC, e DCN) normalmente identificados no estroma de PDAC. Após a avaliação dos tumores primários com RNA-ISH, muitos dos genes identificados, foram encontrados co-localizando em uma sub-população de células na região basal dos ductos pancreáticos malignos. Além disso, observamos que estas células expressam o marcador SV2A neuroendócrino, e o marcador de células estaminais ALDH1A1/2. Em comparação com tumores negativos para SV2, os doentes com tumores SV2 positivos apresentaram níveis mais baixos de CA 19-9 (69% vs. 52%, p = 0,012), tumores maiores (> 4 cm, 23% vs. 10%, p = 0,0430), menor invasão de gânglios linfáticos (69% vs. 86%, p = 0,005) e tumores mais diferenciados (69% vs. 57%, p = 0,047). A presença de SV2A foi associada com uma sobrevida livre de doença mais longa (HR: 0,49 p = 0,009) bem como melhor sobrevida global (HR: 0,54 p = 0,018). Em conjunto, esta informação aponta para dois subtipos diferentes de adenocarcinoma do pâncreas, e estes subtipos co-relacionam estatisticamente com o prognostico de doentes, sendo este subgrupo definido pela presença do clone celular SV2A / ALDH1A1/2 positivo com características neuroendócrinas. No Capítulo 4, a expressão de SV2A no cancro do pâncreas foi validado em linhas celulares primárias. Demonstramos a heterogeneidade do adenocarcinoma do pâncreas de acordo com características clonais neuroendócrinas. Ao comparar as linhas celulares expressando SV2 com linhas celulares negativas, verificamos que as linhas celulares SV2+ eram mais diferenciadas, diferindo de linhas celulares SV2 negativas no que respeita a mutação KRAS, proliferação e a resposta à quimioterapia. No capítulo 5, perguntamo-nos se o clone celular SV2 positivo poderia explicar a resistência a quimioterapia observada em doentes. Observamos um aumento absoluto de clones celulares expressando SV2A, em múltiplas linhas de evidência - doentes, linhas de células primárias e xenotransplantes. Embora, tenhamos sido capazes de demonstrar que o adenocarcinoma do pâncreas é uma doença heterogénea, consideramos que a caracterização genética destes clones celulares expressando SV2A é de elevada importância. Pretendemos colmatar esta limitação com as seguintes estratégias: Após o tratamento com quimioterapia neoadjuvante na nossa coorte, realizamos microdissecação a laser das amostras primarias em parafina, de forma a analisar mutações genéticas observadas no adenocarcinoma pancreático; em segundo lugar, pretendemos determinar consequências de knockdown da expressão de SV2A em nossas linhas celulares seguindo-se o tratamento com gemicitabina para determinação do papel funcional de SV2A; finalmente, uma vez que os nossos esforços anteriores com um promotor - repórter e SmartFlare ™ falharam, o próximo passo será realizar RNA-ISH PrimeFlow™ seguido de FACS e RNA-seq para caracterização deste clone celular. Em conjunto, conseguimos provar com várias linhas de evidência, que o adenocarcinoma pancreático é uma doença heterogénea, definido por um clone de células que expressam SV2A, com características neuroendócrinas. A presença deste clone no tecido de doentes correlaciona-se estatisticamente com o prognostico da doença, incluindo sobrevida livre de doença e sobrevida global. Juntamente com padrões de proliferação e co-expressão de ALDH1A1/2, este clone parece apresentar um comportamento de células estaminais e está associado a resistência a quimioterapia, uma vez que a sua expressão aumenta após agressão química, quer em doentes, quer em linhas de células primárias.----------------------------- ABSTRACT: Currently, the only chance of cure for patients with pancreatic adenocarcinoma is surgical resection, at the beginning of my thesis studies, we asked if the classical clinicopathologic predictors of outcome could be validated in a large cohort of patients with early stage pancreatic cancer and if other clinical predictors could have a role on deciding which patients would benefit from surgery. In chapter 2, we found that up to 30% of patients die within the first year after curative intent surgery for pancreatic adenocarcinoma. We aimed at determining pre-operative factors that would correlate with early mortality following resection for pancreatic cancer using a statistically validated tool, the Charlson-Age Comorbidity Index (CACI). We found that a CACI score greater than 4 was predictive of increased length of stay (p<0.001), post-operative complications (p=0.042), and mortality within 1-year of pancreatic resection (p<0.001). A CACI score of 6 or greater increased 3-fold the odds of death within the first year. Patients with a high CACI score have less than 50% likelihood of being alive 1 year after surgery. In chapter 3 we aimed at identifying a surface protein that correlates with patient’s outcome and distinguishes sub-groups of patients according to their molecular differences and if this protein could be a cancer stem cell marker. The most abundant class of circulating tumor cells identified in our previous work was found to have biphenotypic features of epithelial to mesenchymal transition, enrichment for stem-cell associated genes (ALDH1A1/ALDH1A2 and KLF4), and an overexpression of extracellular matrix genes (Collagens, SPARC, and DCN) normally found in the stromal microenvironment of PDAC primary tumors. Upon evaluation of matched primary tumors with RNA-ISH, many of the genes identified were found to co-localize in a sub-population of cells at the basal region of malignant pancreatic ducts. In addition, these cells expressed the neuroendocrine marker SV2A, and the stem cell marker ALDH1A1/2. Compared to SV2 negative tumors, patients with SV2 positive tumors were more likely to present with lower CA 19-9 (69% vs. 52%, p = 0.012), bigger tumors (size > 4 cm, 23% vs. 10%, p= 0.0430), less nodal involvement (69% vs. 86%, p = 0.005) and lower histologic grade (69% vs. 57%, p = 0.047). The presence of SV2A expressing cells was associated with an improved disease free survival (HR: 0.49 p=0.009) and overall survival (HR: 0.54 p=0.018) and correlated linearly with ALDH1A2. Together, this information points to two different sub-types of pancreatic adenocarcinoma, and these sub-types correlated with patients’ outcome and were defined by the presence of a SV2A/ ALDH1A1/2 expressing clone with neuroendocrine features. In Chapter 4, SV2A expression in cancer was validated in primary cell lines. We were able to demonstrate pancreatic adenocarcinoma heterogeneity according to neuroendocrine clonal features. When comparing SV2 expressing cell lines with SV2 negative cell lines, we found that SV2+ cell lines were more differentiated and differ from SV2 negative cell lines regarding KRAS mutation, proliferation and response to chemotherapy. In Chapter 5 we aimed at determining if this SV2 positive clone could explain chemoresistance observed in patients. We found an absolute increase in SV2A expressing cells, with multiple lines of evidence, in patients, primary cell lines and xenografts. Although, we have been able to show evidence that pancreatic adenocarcinoma is a heterogeneous disease, our findings warrant further investigation. To further characterize SV2A expressing clones after treatment with neoadjuvant chemotherapy in our cohort, we have performed laser capture microdissection of the paraffin embedded tissue in this study and will analyze the tissue for known genetic mutations in pancreatic adenocarcinoma; secondly, we want to know what will happen after knocking down SV2A expression in our cell lines followed by treatment with gemcitabine to determine if SV2A is functionally important; finally, since our previous efforts with a promoter – reporter and SmartFlare™ have failed, we will utilize a novel PrimeFlow™ RNA-ISH assay followed by FACS and RNA sequencing to further characterize this cellular clone. Overall our data proves, with multiple lines of evidence, that pancreatic adenocarcinoma is a heterogeneous disease, defined by a clone of SV2A expressing cells, with neuroendocrine features. The presence of this clone in patients’ tissue correlates with patient’s disease free survival and overall survival. Together with patterns of proliferation and ALDH1A1/2 co-expression, this clone seems to present a stem-cell-like behavior and is associated with chemoresistance, since it increases after chemotherapy, both in patients and primary cell lines.

博士
國立陽明大學
藥理學研究所
99
Oct4 and Nanog are pluripotency genes, but their roles in adult stem cells are unclear. Here, increase in Oct4 and Nanog expression along with increased proliferation and differentiation potential but decreased spontaneous differentiation were observed in early passage (E), hypoxic culture (H) and p21 knockdown (p21KD) mesenchymal stem cells (MSCs) compared to late passage (L), normoxic culture (N) and scrambled shRNA-overexpressed (Scr) MSCs. Knockdown of Oct4 and Nanog in E, H and p21KD MSCs decreased proliferation and differentiation potential, and enhanced spontaneous differentiation, whereas overexpression of Oct4 and Nanog in L, N and Scr MSCs increased proliferation and differentiation potential, and suppressed spontaneous differentiation. Moreover, E, H and p21KD MSCs expressed higher Dnmt1 compared to L, N and Scr MSCs. Using 5 aza-cytidine, the potent DNA demethylating agent, decreased proliferation and differentiation potential, and enhanced spontaneous differentiation in E, H and p21KD MSCs. These data demonstrate the roles of Oct4 and Nanog in maintaining self-renewal and undifferentiated state in MSCs.

碩士
中原大學
生物醫學工程研究所
105
The study used D-galactose (D-gal) treated bone marrow mesenchymal stem cells (MSCs) as an aging model, and aimed to explore the influence of low level light irradiation (LLLI) on anti-senescence of stem cells. We first determined cell viability, activity of senescence-associated- β-galactosidase (SA-β-gal), reactive oxygen species (ROS) and malondialdehyde (MDA) level on D-gal treated MSCs. SA-β-gal activity, ROS and MDA level were found to increase with increasing concentration of D-gal, whereas cell viability decreased. After red (630 nm) and near-infrared (850 nm) light irradiation, MDA concentration and SA-β-gal activity declined, whereas cell viability elevated as compared to the control group. The results suggest that D-gal treatment could induce senescence of MSCs via oxidative stress. LLLI could delay MSCs senescence by reducing oxidative stress. In addition, the effect of LLLI to MSCs senescence gene, p53, p21, p16 expression and osteogenic marker expression was tested. The results showed p53, p21, and p16 senescence gene expression increased when increasing D-gal concentration while it decreased after LLLI. In osteogenic marker expression, increasing D-gal concentration inhibited alkaline phosphatase (ALP) and calcium deposition expression. Although ALP expression was lower than control group, calcium deposition expression was higher than control group after LLLI. Overall, D-gal is able to induce MSCs senescence and reduces MSCs growing and osteogenic differentiation ability. LLLI treatment can effectively delay MSCs senescence and induced MSCs growing and osteogenic differentiation.

博士
國立成功大學
生物科技研究所碩博士班
97
In vivo, postnatal stem cells are usually quiescent and drug resistant; they can be prompted to proliferate when triggered by proper signals. In vitro, however, stem cells propagate quickly and differentiate spontaneously. Therefore, holding stem cells in vitro with reduced growth and multidrug resistance (MDR) phenotype is an appropriate strategy for keeping them in a primitive state. Because hyaluronan has been recognized as a crucial regulator for maintaining the microenvironments termed stem cell niches, we examined whether hyaluronan induces slow cycling and drug resistance in placenta-derived mesenchymal stem cells (PDMSCs) by comparing hyaluronan-coated surface with tissue-culture polystyrene surface. The hyaluronan-coated surface significantly downregulated the proliferation of PDMSCs, more of which were maintained in the G0/G1 phases than were cells on the tissue-culture polystyrene surface. Both PKH-26 labeling and BrdU incorporation assays showed that most PDMSCs grown on a hyaluronan-coated surface duplicated during cultivation indicating that the hyaluronan-coated surface did not inhibit PDMSCs from entering the cell cycle. Mitotic synchronization showed that the G1-phase transit was prolonged in PDMSCs growing on a hyaluronan-coated surface. In addition, increases in p27Kip1 and p130 were the crucial factors that allowed hyaluronan to lengthen the G1 phase. We found that PDMSCs cultured on a tissue-culture polystyrene surface coated with 30 μg/cm2 hyaluronan were more resistant to doxorubicin compared to control PDMSCs. Inhibiting PI3-K/Akt signaling showed that the PI3-K/Akt pathway modulated both P-glycoprotein activity and doxorubicin resistance. In addition, 10 μM verapamil dramatically suppressed the doxorubicin resistance induced by the hyaluronan-coated surface, indicating that P-glycoprotein activity was necessary for MDR. We further demonstraterd that PDMSCs treated with CD44 small interfering RNA (siRNA) and grown on a polystyrene surface coated with 30 μg/cm2 hyaluronan had fewer P-glycoprotein+ cells and lower CD44 expression levels (less than 60% in both cases) compared with PDMSCs not treated with CD44 siRNA and grown on the hyaluronan-coated surface. Moreover, treatment with CD44 siRNA suppressed the hyaluronan-substratum-induced resistance of PDMSCs to doxorubicin. The data provided herein demonstrated that hyaluronan might be a promising candidate for maintaining stem cells in slow-cycling mode by prolonging their G1-phase transit. In addition, we presume that induction of MDR by hyaluronan would hold PDMSCs in a primitive state by providing the capability to extrude molecules required for differentiation. It seems reasonable to suggest that hyaluronan may cause PDMSCs to enter a program of dormancy, the natural state of stem cells consisting with slow cycling and drug resistance. Therefore, our findings may contribute to a deeper understanding of physiological functions of hyaluronan in stem cell research, and that is valuable for the applications in regenerative medicine.

Embryonic stem cells (ES-cells) are an excellent source for generating insulin-producing β-islet cells for potential use in the management of diabetes mellitus. Although many protocols have been developed to promote the differentiation of ESCs into β-islet cells, they are limited in terms of (i) low efficiency of differentiation and (ii) generation of functionally immature β-islet cells with inefficient glucose stimulated insulin secretion (GSIS). The present study is aimed at understanding the differentiation and functional maturity of pancreatic β-islet cell. Earlier in our lab EGFP-expressing transgenic mouse ‘GS-2’ ES-cell line was derived (Singh et al., 2012). Through the embryoid body (EB) formation and differentiation method (day 2 through 21), we obtained spontaneous differentiation of GS-2 ES-cells to three germ-lineage cell types, with the expression of molecular markers of definitive endoderm (DE; sox17) and hepatic lineage (afp). However, we were unable to detect the expression of markers of pancreatic progenitors (pdx1) and β-islet cells (ins2). By improvising the spontaneous differentiation protocol i.e. increasing cell seeding density for EB, we were able to detect expression of pancreatic progenitor marker, pdx-1. However, we were still unable to detect expression of β-islet marker ins1 and ins2. In view of this, we employed an induction protocol by inclusion of laminin, nicotinamide and insulin (Wobus et al., 2006). This resulted in the differentiation of GS-2 ES-cells to DE-like cells by day 14, followed by the appearance of pancreatic progenitor-like clusters by day 21 and significantly, β-islet-like clusters by day 33. This sequential appearance of progenitors and β-islet cells was accompanied by the expression of pdx1, ins1and ins2. However, expression of more important β-islet marker ins1 appeared to be low. Also, we were unable to detect expression of glucose transporter, glut-2 indicating, this model system may not be suitable for testing molecules to improve GSIS or functional maturity. Therefore, we found an alternative model system, Islet Cell Aggregates (ICAs) from day1-2 old neonatal mice that are considered functionally immature or have inefficient GSIS. Sirtuin-1 activator SRT1720 was found as the molecule to improve functional maturity of immature β-islet cells. When functionally immature β-islet cells from day1-2 old neonatal mice were treated with SRT1720 (5μM) for 24 hours, their GSIS improved by 1.5 fold. We envisage that this molecule may be used to improve functional immaturity of PSC-derived functionally-immature β-islet cells

碩士
國立清華大學
科技法律研究所
96
Technological policies have long been influenced by experts. Since the threshold is extremely high for knowledge of technology, it is difficult for the layman to participate in enacting technological policies and to challenge the authority of science experts. Under this kind of frameworks, the common people don’t possess sufficient knowledge to substantially participate in technological decision-making. Therefore, policies determined by experts easily fall outside of the boundaries of democratic politics. By the experience of Missouri Stem Cell Research and Cures Initiative, this thesis introduced the values and functions of “Deliberative Democracy” and its association with technological communication. In contemplation of the experience from Missouri Stem Cell Research and Cures Initiative, this thesis tried to bring in citizen engagement in the procedures of prospective embryo stem cells’ referendum in Taiwan. This would enable the multitude to engage in the public domain of communication and debate, and to make collective decisions approximate to the shared good and in accordance with social justice principles. Hence, it could function to mend the weakening of congressional legislation and further more to enhance the justification of technological policies.

Adult hippocampal neurogenesis entails a continued recruitment of neural precursor cells (NPCs) into active cell cycle and their progressive transition into post-mitotic granule cells. These adult born neurons integrate into the existing circuitry and confer structural plasticity, which aids in key hippocampal functions. For sustained neurogenesis, the cell cycle entry of the NPCs has to be tightly controlled. Environmental cues strongly, and differentially, regulate this checkpoint. Voluntary physical activity represents such an established strong stimulus that results in enhanced proliferation within the neurogenic niche. However, mechanistic insights into the maintenance and regulation of quiescence and the responsiveness of the NPCs to acute physical activity, as a form of adaptive neurogenesis, are yet to be elucidated. In my doctoral studies, we identified redox regulation as a key pathway regulating the cellular state equilibrium. I further explored the role of cellular oxidative stress in the neurogenic course and in adaptive neurogenic responses. Our results show that non-proliferative precursors within the hippocampal dentate gyrus, unlike in other stem cell systems, are marked by high levels of cellular reactive oxygen species (ROS). Using cytometric methodologies, ex vivo bioassays and transcriptional profiling, we revealed that classifying cells based on intracellular ROS content identified functionally defined sub-populations of adult NPCs. We propose that a drop in intracellular ROS content precedes the transition of cellular states, specifically from quiescence to active proliferation. Acute physical activity involves the activation of non- proliferating cells through a transient Nox2-dependent ROS surge in high-ROS, quiescent NPCs. In the absence of Nox2, baseline neurogenesis was unaffected, but the activity- dependent response was abolished. These findings shed new light on the discrete cellular events, which maintain the homeostasis between distinct cellular states of NPCs within the adult murine hippocampus.:Zusammenfassung 3 Summary 4 Acknowledgements 5 Index 8 List of figures 10 List of tables 11 Abbreviations 12 Publications 14 Introduction 15 Adult hippocampal neurogenesis 16 Adult subventricular neurogenesis 21 Methods to study adult neurogenesis 23 Environmental regulation of neurogenesis 26 Redox regulation in a stem cell 29 Working hypothesis 31 Specific aims 31 Materials and methods 32 Mice 34 Physical activity paradigm 35 Thymidine labelling and tissue preparation 35 Fluorescence immunohistochemistry 35 DG and SVZ dissection and dissociation 36 Flow cytometry 36 Gating for ROS classes 36 Neurosphere culture 37 Generation of monolayer culture 37 Inducing quiescence through BMP4 treatment 38 Next Generation sequencing (NGS) 38 RNA extraction 38 Quality control and differential expression 39 Functional enrichment and expression profiles 41 RNA isolation and quantitative RTPCR (qRT-PCR) 43 Ki67 immunochemistry and quantification of in vivo proliferation 45 Quantification and statistical analysis 46 Data and software availability 48 Results 49 Intracellular ROS content functionally delineates subpopulations of neural precursor cells 49 Resolution of ROS profiles of DG and SVZ and neurosphere bioassay 49 Distribution of Nes-GFP cells into different ROS classes 54 Neural precursors of the different ROS classes have distinct molecular profiles 55 Changes in intracellular ROS content precede cell fate changes 65 ROS profiling of other cell types within the DG 70 ROS profiling of Astrocytes and type-1 cells 70 ROS profiling of Doublecortin (Dcx)positive cells of the neurogenic lineage 74 ROS profiling of microglial cells within the DG 77 Resolving the response of Nes-GFP subpopulations to environmental stimulus 78 Nes-GFP+ cells of the hiROS class specifically respond to physical activity 81 Changes in ROS content are not driven by mitochondrial activity 83 In vitro monolayer culture of NPCs as an independent corroboration 86 Discussion 89 The organization of an active stem cell niche with respect to redox content 89 Cytometric classification of cells within the DG 91 Establishing the cellular states of redox defined subsets of Nes-GFP+ adult precursors within the DG 95 Timeline of baseline proliferation within precursors and identifying the subset of precursors responsive to de novo physical activity 97 Monolayer culture to study cellular states and redox regulation 100 Nox2 dependency as a discriminatory feature of adaptive neurogenesis 101 Conclusion 103 References 104 Declarations 122 Anlage 1 122 Anlage 2 124

碩士
國立臺灣大學
健康政策與管理研究所
99
The Clinical appliance of Stem Cells started from 8th century, meanwhile the Research went to a new mile stone since from 19th century. The continues research combine with lots of experts, clinicians, professors, technologists of Clinical Medicine, Basic Medicine, Medical Technology, Pharmacology, Dentistry, Material Engineering, Molecular Biology, BioTechnology, Bio Medicine of different fields from different countries has been developing day after day. Modern Technology of Stem Cells widely use Adult Autologus, Allogeneous, Xenogeneous Stem Cells add the mature technique of HLA-typing and the well understanding of GvHD which caused by Allogeneous, Xenogeneous Stem Cells implantation, More and more large scaled animal or Human Clinical Trials were successfully practiced under using the getting matured technique of Genes transplantation,combined therapy of Stem Cells and or Tissue Engineering of Stem Cells. More and more news were broadcasted in front of people which revealed many diseases which could not be cured under so called Current Medical Care werecured ,getting improved, recovered under the aid of Stem Cells .The heart of people full filled of hope to Stem Cells. We must have the vi combination of reasonable and sufficient laws, rulings ,and policies to make Taiwan as the island of Bio technology while the government of Taiwan develop Bio technology strongly. Whether the IT industry will be replaced by the industry of Bio technology which make Taiwan become the island of Bio technology industry, and it depend on the orientations and policies of the government as the developing of Stem Cells will be listed as the focus and superior or rewarded item of Biotechnology. The Research use the Qualitative Researching methods which select many interviewers belong to certain manufacturing ,government ,and professions to interviewed by the methods of In-depth interview. Finally the conclusion was made according to the analysis of the research. We do the cross comparison and analysis of the interviewers without the classification of their occupation of the research . The generalized conclusion of the research as like the following items: they are (1). If the Stem Cells Research will become one part of Main stremed Medicine? Yes: 83.4 %, No: 0% ,Not sure: 16.6%. (2). Wheather Autologus or Allogeneous Stem Cells will become the Main Streamed Ste, Cells Research? Autologus:50 %, Allogeneous:50%.(3).(3.1).The standard of Taiwan Stem Cells Research ? Has reached the international standard: 0%, Not yet reached the international standard: 100%,Not sure: 0%.(3.2)Does Taiwan need import the advanced Stem Cells technique overseas? Yes:100%,No:0%,Not sure: 0%. (4).(4.1) Concepts to the Umbilical Cord Blood vii Storage in Taiwan? Meaningful:0%, Not too meaningful:100%, Not clear:0%.(4.2).Futuring of the Umbilical Cord Blood storage in Taiwan ? Good futuring: 0%,Not good futuring: 100%, Not clear:0%. (5).Imported International technique of Adult Stem Cells of storage and treatment to Taiwan? Need: 100%,No need:0%,not clear:0%/. (6).What orientation should be adopted to the Stem Cells policies? Opened under certain qualifications: 100%, Policies should be restricted:0%, no comments: 0%. (7).(7.1)Should Taiwan government treat Bio-technique industries as the first priority of the government policies ? Yes: 83.4%, No:0%,not clear: 16.6%. (7.2) Should Taiwan government treat and reward the research of Stem Cells as the first priority of her policies? Yes: 41.55%, No: 8.7%, not sure: 49.8%. (8). Do you agree with adoption Stem Cells treatment to the payment of generalized health insurance in Taiwan? Yes: 83.34, No: 0%, Yes under certain situations: 16.66%. Hopefully, the result can be treated as the main resource to the related officials, decisions makers, certain persons of professions of medical, Bio medical, Stem cells research, medical policies, related fields of industries.

The work deals with looking at the status of the human embryo. The structure of the work is divided into several chapters that deal with a particular topic. The first chapter deals with the historical view of the adoption of a human embryo, in the second chapter is devoted to philosophical adopting the human embryo and view individual philosophers on this issue. The third chapter is devoted to the ethical acceptance of the issue. Here is discussed an ethical perspective on human embryo status and access two main streams which occur in this area. The next part is devoted to the present, both in terms of the ethical-philosophical, and in terms of health. In the medical section includes prenatal communication between mother and child, and research on embryonic stem cells, which is associated with a number of ethical issues.