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Добірка наукової літератури з теми "Embryonic stem cells"

Автор: Grafiati

Опубліковано: 4 червня 2021

Оновлено: 29 липня 2024

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Статті в журналах з теми "Embryonic stem cells"

1

Pera, M. F., B. Reubinoff, and A. Trounson. "Human embryonic stem cells." Journal of Cell Science 113, no. 1 (January 1, 2000): 5–10. http://dx.doi.org/10.1242/jcs.113.1.5.

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Embryonic stem (ES) cells are cells derived from the early embryo that can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent; they share these properties with embryonic germ (EG) cells. Candidate ES and EG cell lines from the human blastocyst and embryonic gonad can differentiate into multiple types of somatic cell. The phenotype of the blastocyst-derived cell lines is very similar to that of monkey ES cells and pluripotent human embryonal carcinoma cells, but differs from that of mouse ES cells or the human germ-cell-derived stem cells. Although our understanding of the control of growth and differentiation of human ES cells is quite limited, it is clear that the development of these cell lines will have a widespread impact on biomedical research.

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2

HOSSEINI, Hamid, and S. MOOSAVI-NEJAD. "1A34 Shock waves effects on embryonic stem cells." Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME 2014.26 (2014): 35–36. http://dx.doi.org/10.1299/jsmebio.2014.26.35.

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3

Challa, Stalin Reddy, and Swathi Goli. "Differentiation of Human Embryonic Stem Cells into Engrafting Myogenic Precursor Cells." Stem cell Research and Therapeutics International 1, no. 1 (April 16, 2019): 01–05. http://dx.doi.org/10.31579/2643-1912/002.

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Degenerative muscle diseases affect muscle tissue integrity and function. Human embryonic stem cells (hESC) are an attractive source of cells to use in regenerative therapies due to their unlimited capacity to divide and ability to specialize into a wide variety of cell types. A practical way to derive therapeutic myogenic stem cells from hESC is lacking. In this study, we demonstrate the development of two serum-free conditions to direct the differentiation of hESC towards a myogenic precursor state. Using TGFß and PI3Kinase inhibitors in combination with bFGF we showed that one week of differentiation is sufficient for hESC to specialize into PAX3+/PAX7+ myogenic precursor cells. These cells also possess the capacity to further differentiate in vitro into more specialized myogenic cells that express MYOD, Myogenin, Desmin and MYHC, and showed engraftment in vivo upon transplantation in immunodeficient mice. Ex vivo myomechanical studies of dystrophic mouse hindlimb muscle showed functional improvement one month post-transplantation. In summary, this study describes a promising system to derive engrafting muscle precursor cells solely using chemical substances in serum-free conditions and without genetic manipulation.

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4

Cezar, Gabriela Gebrin. "Embryonic Stem Cells." International Journal of Pharmaceutical Medicine 20, no. 2 (2006): 107–14. http://dx.doi.org/10.2165/00124363-200620020-00004.

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5

Wagner, Erwin F. "Embryonic stem cells." Current Opinion in Oncology 4 (December 1992): S2—S4. http://dx.doi.org/10.1097/00001622-199212001-00002.

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6

Rippon, H. J., and A. E. Bishop. "Embryonic stem cells." Cell Proliferation 37, no. 1 (February 2004): 23–34. http://dx.doi.org/10.1111/j.1365-2184.2004.00298.x.

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7

Biswas, Atindriya, and Robert Hutchins. "Embryonic Stem Cells." Stem Cells and Development 16, no. 2 (April 2007): 213–22. http://dx.doi.org/10.1089/scd.2006.0081.

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8

Etches, Robert J. "Embryonic stem cells." Lancet Oncology 2, no. 3 (March 2001): 131–32. http://dx.doi.org/10.1016/s1470-2045(00)00252-7.

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9

Hampton, Tracy. "Embryonic Stem Cells." JAMA 297, no. 5 (February 7, 2007): 459. http://dx.doi.org/10.1001/jama.297.5.459-a.

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Morowitz, Harold. "Embryonic stem cells." Complexity 8, no. 3 (January 2003): 10–11. http://dx.doi.org/10.1002/cplx.10080.

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Дисертації з теми "Embryonic stem cells"

1

Nortjé, Nico. "The moral status of embryonic stem cell research in the South African context /." Link to the online version, 2007. http://hdl.handle.net/10019.1/1372.

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2

Harun, Rosliah. "Derivation of trophoblast stem cells from human embryonic stem cells." Thesis, University of Sheffield, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.414643.

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3

Montgomery, Sarah Lynn. "Impedance measurement system for embryonic stem cell and embryoid body cultures." Thesis, Atlanta, Ga. : Georgia Institute of Technology, 2008. http://hdl.handle.net/1853/24661.

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4

Gilner, Jennifer Bushman Kirby Suzanne Lee. "Enrichment of therapeutic hematopoietic stem cell populations from embryonic stem cells." Chapel Hill, N.C. : University of North Carolina at Chapel Hill, 2007. http://dc.lib.unc.edu/u?/etd,1232.

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Thesis (Ph. D.)--University of North Carolina at Chapel Hill, 2007.
Title from electronic title page (viewed Mar. 26, 2008). "... in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Pathology and Laboratory Medicine." Discipline: Pathology and Laboratory Medicine; Department/School: Medicine.

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5

Jurczak, Daniel. "Stemness in human embryonic stem cells." Thesis, University of Skövde, School of Life Sciences, 2009. http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-3509.

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Stem cells are cells that have a unique ability to divide for an indefinite period. Additionally, they can give rise to a plethora of specialized cell types. The advent of high-throughput technologies made it possible to investigate gene expression on a large scale. This enabled scientists to perform comprehensive gene profiling studies of stem cells. Several authors have suggested that there might be a common set of genes that control the stemness of stem cells. In this study, we suggest that ”stemness” genes that are related to ”stemness” characteristics show a statistically significant down-regulation between undifferentiated and differentiated cells. For this we have analyzed microarray data from five different cell lines and compared their global expression profiles. Common down-regulated transcripts among those data sets were de- rived by using a well-established gene expression analysis procedure called Significance Analysis of Microarrays. Since all three data sets were provided by Cellartis AB, the derived list of common transcripts was subsequently compared with an external study. Moreover, we also performed a comparison with down-regulated genes derived from mouse embryonic stem cells. This was done to determine if there is a common set of stemness genes even across distinct species. Re- sults were further evaluated using a comprehensive data-set from a study by Skottman et al. (2005). All results where compared uti- lizing using a range of false discovery rate threshold values and the results were subsequently used for gene ontology term enrichment. GO terms where utilized to functionally annotate and classify those embryonic stem cell transcripts, that were found to be common in all data-sets and identify over-represented biological processes related to those genes.

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6

Nussbaum, Jeannette. "Embryonic stem cells for myocardial infarct repair /." Thesis, Connect to this title online; UW restricted, 2004. http://hdl.handle.net/1773/6312.

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7

Noisa, Parinya. "Characterization of neural progenitor/stem cells derived from human embryonic stem cells." Thesis, Imperial College London, 2010. http://hdl.handle.net/10044/1/5712.

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Human embryonic stem cells (hESCs) are able to proliferate indefinitely without losing their ability to differentiate into multiple cell types of all three germ layers. Due to these fascinating properties, hESCs have promise as a robust cell source for regenerative medicine and as an in vitro model for the study of human development. In my PhD study, I have investigated the neural differentiation process of hESCs using our established protocol, identified characteristics associated with each stage of the differentiation and explored possible signalling pathways underlying these dynamic changes. It was found that neural differentiation of hESCs could be divided into 5 stages according to their morphology, marker expression and differentiation potencies: hESCs, neural initiation, neural epithelium/rosette, neuronal progenitor cells and neural progenitor/stem cells (NPSCs) and 4 of these stages have been studied in more detail. At the neural initiation, hESCs firstly lose TRA-1-81 expression but retain SSEA4 expression. This transient cell population shows several similar properties to the primitive ectoderm. After neural-tube like structure/neural rosette formation, neural progenitor cells appear as typical bipolar structures and exhibit several properties of radial glial cells, including gene expression and pro-neuronal differentiation. The neural progenitor cells are able to grow in culture for a long time in the presence of growth factors bFGF and EGF. However, they gradually lose their bipolar morphology to triangular cell type and become pro-glial upon further differentiation. In addition, the state of neural progenitor and stem cells can be distinguished by their differential response to canonical Notch effector, C protein-binding factor 1. It was also found that delta like1 homolog (DLK-1) is temporally upregulated upon initial neural differentiation, but becomes undetectable after the neural progenitor stage. Overexpression of DLK-1 in NPSCs enhances neuronal differentiation in the presence of serum by blocking BMP and Notch pathways. These results show that neural differentiation of hESCs is a dynamic process in which cells go through sequential changes, and the events are reminiscent of the in vivo neurodevelopment process. Moreover, I have characterized stably transfected nestin-GFP reporter hESC lines and found that the cell lines maintained the features of hESCs and the expression of GFP is restricted to the neural lineage after differentiation. Therefore, these reporter lines will be useful for the study of factors that regulate neural differentiation and for the enrichment of neural progenitors from other lineages. Taken together, this study has demonstrated that hESCs are a good in vitro model to study the mechanisms and pathways that are involved in neural differentiation. The availability of hESCs allows us to explore previously inaccessible processes that occur during human embryogenesis, such as gastrulation and neurogenesis.

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Bigdeli, Narmin. "Derivation, characterization and differentiation of feeder-free human embryonic stem cells /." Göteborg : Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine at Sahlgrenska Academy, University of Gothenburg, 2010. http://hdl.handle.net/2077/22353.

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9

Lu, Xibin, and 盧希彬. "Quantitative characterization of mouse embryonic stem cell state transition." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2014. http://hdl.handle.net/10722/208049.

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Bowles, K. M. "Generation of haematopoietic cells from human embryonic stem cells." Thesis, University of Cambridge, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.596829.

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Анотація:

Culture of hESCs on murine stromal layers or in stromal free conditions as embryoid bodies results in low levels of haematopoietic cells. Here it is demonstrated that overexpression of the transcription factor HOXB4 considerably augments haematopoetic development of hESCs. Stable HOXB4 expressing hESC clones were generated by lipofection and could be maintained in the undifferentiated state for prolonged passages. Moreover, differentiation of hESCs as embryoid bodies in serum containing medium without the use of additional of cytokines led to sequential expansion of first erythroid then myeloid and monocytic progenitors. These cells retained the capacity to develop into formed blood elements during in vitro culture. Consistent with the development of committed haematopoietic cells the expression of transcription factors known to be critical for haematopoietic development was observed. The successful use of enforced gene expression to promote the differentiation of hESCs into a terminally differentiated tissue is thus demonstrated, thereby revealing an important role for HOXB4 in supporting their in vitro development along the haematopoietic pathway. Once a method for producing significant numbers of haematopoietic cells from hESCs in vitro was established, hESCs were used to study the role of stem cell leukaemia gene (SCL) in human blood and endothelial development. hESC lines were generated in which the expression of SCL, under control of an enhancer previously shown in mice to target expression to blood and endothelial progenitors, was evident through green fluorescent protein (GFP) expression. The enhancer directed GFP expression inhuman K562 cells and some differentiated progeny of HOXB4 transfected hESCs. However a more thorough assessment of GFP positive cells was hindered by problems with transgene silencing.

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Книги з теми "Embryonic stem cells"

1

Irina, Klimanskaya, and Lanza Robert, eds. Embryonic stem cells. Amsterdam: Elsevier Academic Press, 2006.

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2

W, Masters J. R., Palsson Bernhard, and Thomson James A. Dr, eds. Embryonic stem cells. Dordrecht: Springer, 2007.

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3

Turksen, Kursad. Embryonic Stem Cells. New Jersey: Humana Press, 2001. http://dx.doi.org/10.1385/1592592414.

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4

V, Greer Erik, ed. Embryonic stem cell research. New York: Nova Science Publishers, 2006.

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5

Chiu, Arlene, and Mahendra S. Rao. Human Embryonic Stem Cells. New Jersey: Humana Press, 2003. http://dx.doi.org/10.1385/1592594239.

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6

Sullivan, Stephen, Chad A. Cowan, and Kevin Eggan, eds. Human Embryonic Stem Cells. Chichester, UK: John Wiley & Sons, Ltd, 2007. http://dx.doi.org/10.1002/9780470511619.

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Chiu, Arlene Y., and Mahendra S. Rao, eds. Human Embryonic Stem Cells. Totowa, NJ: Humana Press, 2003. http://dx.doi.org/10.1007/978-1-59259-423-8.

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8

S, Odorico Jon, Zhang S. -C, and Pedersen Roger A, eds. Human embryonic stem cells. Abingdon, Oxon, UK: Garland Science/BIOS Scientific Publishers, 2006.

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9

Kiessling, Ann A. Human embryonic stem cells. 2nd ed. Sudbury, Mass: Jones and Bartlett Publishers, 2007.

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10

1951-, Anderson Scott, ed. Human embryonic stem cells. 2nd ed. Sudbury, Mass: Jones and Bartlett Publishers, 2007.

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Частини книг з теми "Embryonic stem cells"

1

Jones, D. Gareth. "Stem Cells: Embryonic." In Encyclopedia of Global Bioethics, 2690–96. Cham: Springer International Publishing, 2016. http://dx.doi.org/10.1007/978-3-319-09483-0_401.

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2

Jones, D. Gareth. "Stem Cells: Embryonic." In Encyclopedia of Global Bioethics, 1–8. Cham: Springer International Publishing, 2014. http://dx.doi.org/10.1007/978-3-319-05544-2_401-1.

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3

Schwab, Manfred. "Embryonic Stem Cells." In Encyclopedia of Cancer, 1. Berlin, Heidelberg: Springer Berlin Heidelberg, 2015. http://dx.doi.org/10.1007/978-3-642-27841-9_1861-2.

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Lewis, Philip, Edina Silajdžić, Daniel R. Brison, and Susan J. Kimber. "Embryonic Stem Cells." In Cell Engineering and Regeneration, 315–65. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-319-08831-0_19.

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Lewis, Philip, Edina Silajdžić, Daniel R. Brison, and Susan J. Kimber. "Embryonic Stem Cells." In Cell Engineering and Regeneration, 1–51. Cham: Springer International Publishing, 2017. http://dx.doi.org/10.1007/978-3-319-37076-7_19-1.

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6

Eiges, Rachel, Naomi Zak, Benjamin E. Reubinoff, and Charles S. Irving. "Embryonic Stem Cells." In Stem Cells in Regenerative Medicine, 447–86. Chichester, UK: John Wiley & Sons, Ltd, 2016. http://dx.doi.org/10.1002/9781118846193.ch25.

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7

Browning, Victoria L., and Jon S. Odorico. "Embryonic Stem Cells." In Stem Cells in Endocrinology, 3–22. Totowa, NJ: Humana Press, 2005. http://dx.doi.org/10.1385/1-59259-900-1:003.

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8

ten Have, Henk, and Maria do Céu Patrão Neves. "Stem Cells, Embryonic." In Dictionary of Global Bioethics, 967. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-54161-3_477.

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9

Draper, Jonathan S., Harry Moore, and Peter W. Andrews. "Embryonal Carcinoma Cells." In Human Embryonic Stem Cells, 63–87. Totowa, NJ: Humana Press, 2003. http://dx.doi.org/10.1007/978-1-59259-423-8_4.

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10

Friedel, Roland H. "Targeting Embryonic Stem Cells." In Transgenesis Techniques, 185–97. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-60327-019-9_12.

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Тези доповідей конференцій з теми "Embryonic stem cells"

1

Shu, K., H. Thatte, and M. Spector. "Chondrogenic differentiation of adult mesenchymal stem cells and embryonic stem cells." In 2009 IEEE 35th Annual Northeast Bioengineering Conference. IEEE, 2009. http://dx.doi.org/10.1109/nebc.2009.4967739.

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2

Singh, Ankur, Shalu Suri, Ted T. Lee, Jamie M. Chilton, Steve L. Stice, Hang Lu, Todd C. McDevitt, and Andrés J. Garcia. "Adhesive Signature-Based, Label-Free Isolation of Human Pluripotent Stem Cells." In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80044.

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Анотація:

Generation of human induced pluripotent stem cells (hiPSCs) from fibroblasts and other somatic cells represents a highly promising strategy to produce auto- and allo-genic cell sources for therapeutic approaches as well as novel models of human development and disease1. Reprogramming protocols involve transduction of the Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc into the parental somatic cells, followed by culturing the transduced cells on mouse embryonic fibroblast (MEF) or human fibroblast feeder layers, and subsequent mechanical dissociation of pluripotent cell-like colonies for propagation on feeder layers1, 2. The presence of residual parental and feeder-layer cells introduces experimental variability, pathogenic contamination, and promotes immunogenicity3. Similar to human embryonic stem cells (hESCs), reprogrammed hiPSCs suffer from the unavoidable problem of spontaneous differentiation due to sub-optimal feeder cultures4, growth factors5, and the feeder-free substrate6. Spontaneously differentiated (SD)-hiPSCs display reduced pluripotency and often contaminate hiPSC cultures, resulting in overgrowth of cultures and compromising the quality of residual pluripotent stem cells5. Therefore, the ability to rapidly and efficiently isolate undifferentiated hiPSCs from the parental somatic cells, feeder-layer cells, and spontaneously differentiated cells is a crucial step that remains a bottleneck in all human pluripotent stem cell research.

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3

Sargent, Carolyn Y., Luke A. Hiatt, Sandhya Anantharaman, Eric Berson, and Todd C. McDevitt. "Cardiogenesis of Embryonic Stem Cells is Modulated by Hydrodynamic Mixing Conditions." In ASME 2008 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2008. http://dx.doi.org/10.1115/sbc2008-193129.

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Embryonic stem cells (ESCs) have the potential to differentiate into all somatic cell types and are uniquely capable of differentiating into functional cardiomyocytes; however, to effectively use ESCs for cell-based therapies to regenerate viable myocardial tissue, an improved understanding of mechanisms regulating differentiation is necessary. Currently, application of exogenous factors is commonly attempted to direct stem cell differentiation; however, progression towards controlling multiple environmental factors, including biochemical and mechanical stimuli, may result in increased differentiation efficiency for clinical applications. Additionally, current methods of ESC differentiation to cardiomyocytes are labor-intensive and produce relatively few cardiomyocytes based on initial ESC densities. Rotary suspension culture to produce embryoid bodies (EBs) has been shown to yield greater numbers of differentiating ESCs than static suspension cultures [1]. Thus, the objective of this study was to examine how the hydrodynamic mixing conditions imposed by rotary orbital culture modulate cardiomyocyte differentiation.

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4

Melvin, Tracy, Nicolas Perney, Peter Horak, Neil Hanley, and James Hughes. "Optical Classification of Human Embryonic Stem Cells." In Optical Trapping Applications. Washington, D.C.: OSA, 2013. http://dx.doi.org/10.1364/ota.2013.tt1d.6.

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5

Savchenkova, I. P., E. A. Savchenkova, and Yu A. Osipova. "Murine embryonic stem cells with equine genes." In PROCEEDINGS OF THE II INTERNATIONAL CONFERENCE ON ADVANCES IN MATERIALS, SYSTEMS AND TECHNOLOGIES: (CAMSTech-II 2021). AIP Publishing, 2022. http://dx.doi.org/10.1063/5.0092606.

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6

Pillarisetti, A., H. Ladjal, A. Ferreira, C. Keefer, and J. P. Desai. "Mechanical characterization of mouse embryonic stem cells." In 2009 Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE, 2009. http://dx.doi.org/10.1109/iembs.2009.5333954.

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Chen, Weiqiang, Luis G. Villa-Diaz, Yubing Sun, Shinuo Weng, Jin Koo Kim, Paul H. Krebsbach, and Jianping Fu. "Nanotopography Directs Fate of Human Embryonic Stem Cells." In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80222.

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Анотація:

Human embryonic stem cells (hESCs) have remarkable potentials for breakthroughs in future cell-based therapeutics owing to their self-renewal capability and pluripotency [1–2]. However, their intrinsic mechanosensitivity to biophysical signals from the local cellular microenvironment is not well characterized [3–4]. In this work, we introduced a simple, yet precise, microfabrication strategy for accurate control and patterning of local nanoroughness on glass surfaces using photolithography and reactive ion etching (RIE). Our results demonstrated that nanoscale topological features could provide a potent regulatory signal over a diverse array of hESC behaviors, including their morphology, adhesion, proliferation and clonal expansion, and differentiation.

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Li, Lulu, Rene Schloss, Noshir Langrana, and Martin Yarmush. "Effects of Encapsulation Microenvironment on Embryonic Stem Cell Differentiation." In ASME 2008 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2008. http://dx.doi.org/10.1115/sbc2008-192587.

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Pluripotent embryonic stem cells represent a promising renewable cell source to generate a variety of differentiated cell types. Although many investigators have described techniques to effectively differentiate stem cells into different mature cell lineages, their practicality is limited by the absence of large scale processing consideration and low yields of differentiated cells. Previously we have established a murine embryonic stem cell alginate-poly-l-lysine microencapsulation differentiation system. The three-dimensional alginate microenvironment maintains cell viability, is conducive to ES cell differentiation to hepatocyte lineage cells, and maintains differentiated cellular function. In the present work, we demonstrate that hepatocyte differentiation is mediated by cell-cell aggregation in the encapsulation microenvironment. Both cell aggregation and hepatocyte functions, such as urea and albumin secretion, as well as increased expression of cytokaratin 18 and cyp4507a, occur concomitantly with surface E-cadherin expression. Furthermore, by incorporating soluble inducers, such as retinoic acid, into the permeable microcapsule system, we demonstrate decreased cell aggregation and enhanced neuronal lineage differentiation with the expression of various neuronal specific markers, including neurofilament, A2B5, O1 and GFAP. Therefore, as a result of capsule parameter and microenvironment manipulation, we are capable of targeting cellular differentiation to both endodermal and ectodermal cell lineages.

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Becker, Sven, Stefan Liebner, Werner Seeger, Heinrich Sauer, Wolfgang Clauss, and Robert Voswinckel. "Embryonic Stem Cells As Source For Endothelial Cells For Cell-based Therapies." In American Thoracic Society 2010 International Conference, May 14-19, 2010 • New Orleans. American Thoracic Society, 2010. http://dx.doi.org/10.1164/ajrccm-conference.2010.181.1_meetingabstracts.a4900.

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10

Hu, Yue, Shaohui Pan, Jing Wang, Shanshan Zhang, Long Wang, and Jinlian Hua. "Derivation of oocyte-like cells from mouse embryonic stem cells." In 2010 3rd International Conference on Biomedical Engineering and Informatics (BMEI). IEEE, 2010. http://dx.doi.org/10.1109/bmei.2010.5639615.

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Звіти організацій з теми "Embryonic stem cells"

1

Zuckerman, Kenneth S. Reparative Medicine: Production of Erythrocytes & Platelets from Human Embryonic Stem Cells. Fort Belvoir, VA: Defense Technical Information Center, October 2012. http://dx.doi.org/10.21236/ada566171.

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2

Wong, E. A., and Z. Uni. Modulating intestinal cellular maturation and differentiation in broilers by in ovo feeding. Israel: United States-Israel Binational Agricultural Research and Development Fund, 2018. http://dx.doi.org/10.32747/2018.8134161.bard.

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Анотація:

Mortality in a broiler flock will typically range from 4-5% of the population over the course of 6- 7 weeks in the U.S. and 7-8% of the population in Israel. Suboptimal intestinal maturation and functionality are one of the major factors that contribute to early age mortality and hinder flock body weight uniformity. The development of absorptive and secretory functions is orchestrated by differentiation of cells that arise from stem cells. Supplying compounds by in ovo feeding (IOF) during late embryogenesis provides nutrients that may change the dynamics of stem cell differentiation. We hypothesize that the introduction of specific nutrients or probiotics to the late embryonic chick via IOF will result in an acceleration of the maturation of the small intestine as measured by villus/crypt morphology and the number and distribution of absorptive and secretory cells. A chick that can absorb nutrients more efficiently by increasing the number of cells expressing nutrient transporters and resist enteric pathogens by increasing the number of cells expressing mucin and host defense peptides will be healthier at hatch. This chick may have less need for antibiotics and may show reduced early mortality. The objectives of this proposal are to: 1) develop a model for the development of putative stem cells and absorptive/secretory cells in the small intestine of the late embryonic and early post hatch broiler. 2) determine the ability of IOF of nutrients to modulate the population of differentiated cells in the intestine. 3) determine the ability of IOF of probiotics to modulate the population of differentiated cells in the intestine. 4) reduce early mortality and increase body weight uniformity by IOF of selected nutrients or probiotics. This proposal combines the IOF expertise of Zehava Uni (Hebrew University) with the RNAscope in situ hybridization technique of Eric Wong (Virginia Tech). Previous studies using quantitative PCR to examine expression of genes in the intestine were unable to identify specific cells expressing these genes. RNAscope allows the ability to identify putative stem, absorptive and secretory cells in the small intestine. Thus, we will be able to investigate the effect of IOF on the presence of intestinal absorptive and secretory cells at the cellular level. Understanding the mechanisms for intestinal development and function are key to maintaining peak growth and health of chickens and thus would be of great economic benefit to the poultry industry.

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Wahl, Geoffrey M. A Novel Strategy for Isolation, Molecular and Functional Characterization of Embryonic Mammary Stem Cells Using Molecular Genetics and Microfluidic Sorting. Fort Belvoir, VA: Defense Technical Information Center, June 2008. http://dx.doi.org/10.21236/ada488861.

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4

Funkenstein, Bruria, and Cunming Duan. GH-IGF Axis in Sparus aurata: Possible Applications to Genetic Selection. United States Department of Agriculture, November 2000. http://dx.doi.org/10.32747/2000.7580665.bard.

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Many factors affect growth rate in fish: environmental, nutritional, genetics and endogenous (physiological) factors. Endogenous control of growth is very complex and many hormone systems are involved. Nevertheless, it is well accepted that growth hormone (GH) plays a major role in stimulating somatic growth. Although it is now clear that most, if not all, components of the GH-IGF axis exist in fish, we are still far from understanding how fish grow. In our project we used as the experimental system a marine fish, the gilthead sea bream (Sparus aurata), which inhabits lagoons along the Mediterranean and Atlantic coasts of Europe, and represents one of the most important fish species used in the mariculture industry in the Mediterranean region, including Israel. Production of Sparus is rapidly growing, however, in order for this production to stay competitive, the farming of this fish species has to intensify and become more efficient. One drawback, still, in Sparus extensive culture is that it grows relatively slow. In addition, it is now clear that growth and reproduction are physiological interrelated processes that affect each other. In particular sexual maturation (puberty) is known to be closely related to growth rate in fish as it is in mammals, indicating interactions between the somatotropic and gonadotropic axes. The goal of our project was to try to identify the rate-limiting components(s) in Sparus aurata GH-IGF system which might explain its slow growth by studying the ontogeny of growth-related genes: GH, GH receptor, IGF-I, IGF-II, IGF receptor, IGF-binding proteins (IGFBPs) and Pit-1 during early stages of development of Sparus aurata larvae from slow and fast growing lines. Our project was a continuation of a previous BARD project and could be divided into five major parts: i) obtaining additional tools to those obtained in the previous project that are necessary to carry out the developmental study; ii) the developmental expression of growth-related genes and their cellular localization; iii) tissue-specific expression and effect of GH on expression of growth-related genes; iv) possible relationship between GH gene structure, growth rate and genetic selection; v) the possible role of the IGF system in gonadal development. The major findings of our research can be summarized as follows: 1) The cDNAs (complete or partial) coding for Sparus IGFBP-2, GH receptor and Pit-1 were cloned. Sequence comparison reveals that the primary structure of IGFBP-2 protein is 43-49% identical to that of zebrafish and other vertebrates. Intensive efforts resulted in cloning a fragment of 138 nucleotides, coding for 46 amino acids in the proximal end of the intracellular domain of GH receptor. This is the first fish GH receptor cDNA that had been cloned to date. The cloned fragment will enable us to complete the GH - receptor cloning. 2) IGF-I, IGF-II, IGFBP-2, and IGF receptor transcripts were detected by RT-PCR method throughout development in unfertilized eggs, embryos, and larvae suggesting that these mRNAs are products of both the maternal and the embryonic genomes. Preliminary RT-PCR analysis suggest that GH receptor transcript is present in post-hatching larvae already on day 1. 3) IGF-1R transcripts were detected in all tissues tested by RT-PCR with highest levels in gill cartilage, skin, kidney, heart, pyloric caeca, and brain. Northern blot analysis detected IGF receptor only in gonads, brain and gill cartilage but not in muscle; GH increased slightly brain and gill cartilage IGF-1R mRNA levels. 4) IGFBP-2 transcript were detected only in liver and gonads, when analyzed by Northern blots; RT-PCR analysis revealed expression in all tissues studied, with the highest levels found in liver, skin, gonad and pyloric caeca. 5) Expression of IGF-I, IGF-II, IGF-1R and IGFBP-2 was analyzed during gonadal development. High levels of IGF-I and IGFBP-2 expression were found in bisexual young gonads, which decreased during gonadal development. Regardless of maturational stage, IGF-II levels were higher than those of IGF-L 6) The GH gene was cloned and its structure was characterized. It contains minisatellites of tandem repeats in the first and third introns that result in high level of genetic polymorphism. 7) Analysis of the presence of IGF-I and two types of IGF receptor by immunohistochemistry revealed tissue- and stage-specific expression during larval development. Immunohistochemistry also showed that IGF-I and its receptors are present in both testicular and ovarian cells. Although at this stage we are not able to pinpoint which is the rate-limiting step causing the slow growth of Sparus aurata, our project (together with the previous BARD) yielded a great number of experimental tools both DNA probes and antibodies that will enable further studies on the factors regulating growth in Sparus aurata. Our expression studies and cellular localization shed new light on the tissue and developmental expression of growth-related genes in fish.

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