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Статті в журналах з теми "Nerve tissue Cultures and culture media"
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Pires, Liliana R., Daniela N. Rocha, Luigi Ambrosio, and Ana Paula Pêgo. "The role of the surface on microglia function: implications for central nervous system tissue engineering." Journal of The Royal Society Interface 12, no. 103 (February 2015): 20141224. http://dx.doi.org/10.1098/rsif.2014.1224.
Повний текст джерелаАнотація:
In tissue engineering, it is well accepted that a scaffold surface has a decisive impact on cell behaviour. Here we focused on microglia—the resident immune cells of the central nervous system (CNS)—and on their response to poly(trimethylene carbonate-co-ε-caprolactone) (P(TMC-CL)) fibrous and flat surfaces obtained by electrospinning and solvent cast, respectively. This study aims to provide cues for the design of instructive surfaces that can contribute to the challenging process of CNS regeneration. Cell morphology was evidently affected by the substrate, mirroring the surface main features. Cells cultured on flat substrates presented a round shape, while cells with elongated processes were observed on the electrospun fibres. A higher concentration of the pro-inflammatory cytokine tumour necrosis factor-α was detected in culture media from microglia on fibres. Still, astrogliosis is not exacerbated when astrocytes are cultured in the presence of microglia-conditioned media obtained from cultures in contact with either substrate. Furthermore, a significant percentage of microglia was found to participate in the process of myelin phagocytosis, with the formation of multinucleated giant cells being observed only on films. Altogether, the results presented suggest that microglia in contact with the tested substrates may contribute to the regeneration process, putting forward P(TMC-CL) substrates as supporting matrices for nerve regeneration.
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Liubich, L. D., L. P. Staino, D. M. Egorova, T. D. Skaterna, and E. G. Pedachenko. "Effect of various origins conditioned media on the migration of neural cells in vitro." Fiziolohichnyĭ zhurnal 68, no. 2 (March 11, 2022): 36–50. http://dx.doi.org/10.15407/fz68.02.036.
Повний текст джерелаАнотація:
An important direction in the development of the latest technologies for the restoration of damaged central nervous system is the use of stem/progenitor cells (SPCs), mainly neurogenic SPCs (NSPCs) and mesenchymal multipotent stromal cells (MMSCs). One of the main mechanisms of SPCs action is indirect paracrine effects due to the ability to produce a wide range of biologically active signaling molecules (secretome). The study of regenerative effects of conditioned media (CM) of NSPCs and MMSCs as a source of their secretome seems to be actual and potentially beneficial. The aim of the study is to compare the impact of CM from 24-h cultures of fetal neurogenic cells (NCs (E14), as a source of NSPCs) and adiposederived mononuclear cells (AMCs as a source of MMSCs) on migration capacity of rat neural cells in vitro. AMCs-CM were obtained from 24-h cultures with prevalence of CD105+ cells and ability upon further cultivation to form “spheroids” and potency to differentiate into different cell types. NCs-CM were obtained from 24-h cultures with prevalence of Nestin+ cells and ability upon further cultivation to form “neurospheres” and potency to differentiate into astrocytes (GFAP+) and neurons (β-Tubulin III+). Rat fetal neural cells (E14) were cultured to achieve a confluent monolayer with basic cellular elements of nervous tissue (5-7th day), which was dissected with forming a transection site and DMEM with 10% fetal calf serum (control) or 0.1-0.3 mg/ml (by total protein amount) of NCs-CM or AMCs-CM were added. In control cultures of rat neural cells partial overgrowth of the dissected area of the monolayer was observed due to the migration of cells, formation of a network of processes and intercellular contacts; reaching 13.2% (4th day) – 23.2% (8th day) of its full length. The overgrown area increased after addition of CM: NCsCM – 3 times (0.1-0.2 mg/ml) and 3-4 times (0.3 mg/ml, 4th-8th day), reaching 70.5% of full length of the transection site; AMCs-CM – 1.5 times (0.1-0.2 mg/ml) and 4-7 times (0.3 mg/ml, 4th-8th day), reaching 97.4-100% of full length of the transection site. The addition of NCs CM and AMCs CM resulted in β-catenin translocation into nucleus of cells in rat neural cell cultures, which correlated with the overgrowth of the transection zone. NCs-CM as well as AMCs-CM in dose-dependent manner stimulate migration processes in culture of rat neural cells, obviously, involving β-catenin signaling pathway, contributing to overgrowing of the dissected area (reparation of a mechanical defect). NCs-CM and AMCs-CM are a source of signaling molecules that modulate the microenvironment and activate endogenous repair mechanisms in culture (in vitro model of nerve tissue regeneration).
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Saito, Shigeru, Inas Radwan, Hideaki Obata, Kenichiro Takahashi, and Fumio Goto. "Direct Neurotoxicity of Tetracaine on Growth Cones and Neurites of Growing Neurons In Vitro." Anesthesiology 95, no. 3 (September 1, 2001): 726–33. http://dx.doi.org/10.1097/00000542-200109000-00027.
Повний текст джерелаАнотація:
Background Local anesthetics have direct neurotoxicity on neurons. However, precise morphologic changes induced by the direct application of local anesthetics to neurons have not yet been fully understood. Also, despite the fact that local anesthetics are sometimes applied to the sites where peripheral nerves may be regenerating after injury, the effects of local anesthetics on growing or regenerating neurons have never been studied. Methods Three different neuronal tissues (dorsal root ganglion, retinal ganglion cell layer, and sympathetic ganglion chain) were isolated from an age-matched chick embryo and cultured for 20 h. Effects of tetracaine were examined microscopically and by a quantitative morphologic assay, growth cone collapse assay. Results Tetracaine induced growth cone collapse and neurite destruction. Three neuronal tissues showed significantly different dose-response, both at 60 min and at 24 h after the application of tetracaine (P < 0.01). The ED50 values (mean +/- SD) at 60 min were 1.53+/-1.05 mM in dorsal root ganglion, 0.15+/-0.05 mM in retinal, and 0.06+/-0.02 mM in sympathetic ganglion chain cultures. The ED50 values at 24 h were 0.43+/-0.15 mM in dorsal root ganglion, 0.07+/-0.03 mM in retinal, and 0.02+/-0.01 mM in sympathetic ganglion chain cultures. Concentration of nerve growth factor in the culture media did not influence the ED50 values. The growth cone collapsing effect was partially reversible in dorsal root ganglion and retinal neurons. However, in the sympathetic ganglion culture, no reversibility was observed after exposure to 1 mM tetracaine for 10 or for 60 min. Bupivacaine had similar neurotoxicity to the three types of growing neurons. (The ED50 values at 60 min were 2.32+/-0.50 mM in dorsal root ganglion, 0.96+/-0.16 mM in retinal, and 0.18+/-0.05 mM in sympathetic ganglion chain cultures. The ED50 values at 24 h were 0.34+/-0.09 mM in dorsal root ganglion, 0.21+/-0.06 mM in retinal, and 0.45+/-0.10 mM in sympathetic ganglion chain cultures.) Conclusions Short-term exposure to tetracaine produced irreversible changes in growing neurons. Growth cones were quickly affected, and neurites degenerated subsequently. Sensitivity varied with neuronal type and was not influenced by the concentration of nerve growth factor. Because a similar phenomenon was observed after exposure to bupivacaine, the toxicity to growing neurons may not be unique to tetracaine.
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Walum, Erik, Elisabeth Hansson, and Alan L. Harvey. "In Vitro Testing of Neurotoxicity." Alternatives to Laboratory Animals 18, no. 1_part_1 (November 1990): 153–79. http://dx.doi.org/10.1177/026119299001800118.1.
Повний текст джерелаАнотація:
Many of the toxic compounds that are at large in the environment represent a risk to our neuronal functions. Chemicals may have a direct or indirect effect on the nervous system and they may interfere with general biochemical properties or specific neuronal structures and processes. In this review, a brief presentation of the major neurotoxicological targets is given, together with a discussion of some aspects of the use of different in vitro models for screening purposes and mechanistic studies. It is believed that in vitro methods offer special opportunities for the development of new neurotoxicological assays, and that this development will mainly involve cultured model systems. Therefore, a presentation of nerve and glia tissue culture methods is given, followed by an overview of how information on the action of mercury and mercurials, excitotoxins and acrylamide has been obtained through the use of cultured cell models. It is concluded that the developmental potential in cell neurotoxicology lies within the areas of separation and identification of cells representative for different structures in the nervous system, co-cultivation of different cell types, in vivo/in vitro (ex vivo) procedures, chemically defined media, metabolic competent cultures of human cells and improved physiological conditions for cultivation and exposure.
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Zachos, Nicholas C. "Abstract IA008: Modeling immune-epithelial interactions in health and disease using human intestinal enteroid co-cultures." Cancer Research 82, no. 23_Supplement_1 (December 1, 2022): IA008. http://dx.doi.org/10.1158/1538-7445.crc22-ia008.
Повний текст джерелаАнотація:
Abstract Human intestinal enteroids derived from Lgr5+ actively dividing adult stem cells offer a relevant ex vivo system to study biological processes of the human small intestine and colon. While these primary cultures recreate cellular and functional features of the intestinal epithelium of the small intestine (enteroids) or colon (colonoids), they are limited by the lack of associated cell types, including immunological cells, nerves, stromal cells that help maintain tissue homeostasis and respond to external challenges. In the gut, innate immune cells interact with the epithelium by conducting lumial surveillance, support barrier function, and deploy effector functions in response to injury/damage. We have established a co-culture system of enteroid/colonoid monolayers and underlying macrophages and polymorphonuclear neutrophils to recapitulate the cellular framework of the human intestinal epithelial niche. Human intestinal enteroids can generated from biopsies or resected tissue from any segment of the human intestine/colon and maintained in long-term cultures in a basement membrane-rich matrix supplemented with growth factor-conditioned media necessary to maintain intestinal stem cell homeostasis. Immune cells are isolated from fresh human whole blood or frozen peripheral blood mononuclear cells (PBMC) and co-cultured with human intestinal enteroids/colonoids grown as confluent monolayers on permeable supports (e.g., Transwells). This configuration allows for controlled access to the apical side of the epithelium to mimic the gut luminal environment as well as the basolateral side occupied by immune cells to mimic the mucosal environment. Enteroid-immune co-cultures enable multiple outcome measures, including transepithelial resistance, production of cytokines/chemokines, phenotypic analysis of immune cells, tissue immunofluorescence imaging, protein or mRNA expression, antigen or microbe uptake, and other cellular functions. Citation Format: Nicholas C. Zachos. Modeling immune-epithelial interactions in health and disease using human intestinal enteroid co-cultures [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr IA008.
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Lischer, Mirko, Pietro G. di Summa, Carlo M. Oranges, Dirk J. Schaefer, Daniel F. Kalbermatten, Raphael Guzman, and Srinivas Madduri. "Human platelet lysate stimulated adipose stem cells exhibit strong neurotrophic potency for nerve tissue engineering applications." Regenerative Medicine 15, no. 3 (March 2020): 1399–408. http://dx.doi.org/10.2217/rme-2020-0031.
Повний текст джерелаАнотація:
Aim: We investigated a potential strategy involving human platelet lysate (HPL) as a media additive for enhancing the neurotrophic potency of human adipose stem cells (ASC). Materials & methods: Dorsal root ganglion explants, ASC and Schwann cells were used for in vitro axonal outgrowth experiments. Results: Remarkably, HPL-supplemented ASC promoted robust axonal outgrowth, in other words, four-times higher than fetal bovine serum-supplemented ASC and even matched to the level of Schwann cells. Further, analysis of regime of growth medium additive supplementation revealed the critical play of HPL in dorsal root ganglion and stem cells co-culture system for mounting effective axonal growth response. Conclusion: HPL supplementation significantly improved the neurotrophic potency of ASC as evidenced by the robust axonal outgrowth; these findings hold significance for nerve tissue engineering applications.
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Meftahpour, Vafa, Somaiyeh Malekghasemi, Amir Baghbanzadeh, Ali Aghebati-Maleki, Ramin Pourakbari, Ali Fotouhi, and Leili Aghebati-Maleki. "Platelet lysate: a promising candidate in regenerative medicine." Regenerative Medicine 16, no. 1 (January 2021): 71–85. http://dx.doi.org/10.2217/rme-2020-0065.
Повний текст джерелаАнотація:
Human platelet lysate has attracted much interest from many researchers as it is growth-factor rich for cell expansion, which is employed as a new therapeutic strategy. Not only are human platelet lysates used for cell therapy, but they are also used for the completion of basal media in mesenchymal stem cell cultures. Due to the presence of a large number of growth factors, platelet lysates have potential roles in wound healing, treatment of ocular graft-versus-host disease, osteoarthritis, Parkinson’s disease, tendon regeneration, infertility, androgenetic alopecia, nerve repair and regenerative tissue, such as bone regeneration. In this review, we summarize that platelet lysates could be valuable candidates for the treatment of a variety of diseases in regenerative medicine.
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Хасанов, Р. Р., А. А. Гумеров, К. Х. Шефер, and Л. М. Вессель. "A method for culturing of enteric nervous system cells suitable for tissue engineering of the intestine." ZHurnal «Patologicheskaia fiziologiia i eksperimental`naia terapiia», no. 2() (May 27, 2019): 132–41. http://dx.doi.org/10.25557/0031-2991.2019.02.132-141.
Повний текст джерелаАнотація:
Лечение синдрома короткой кишки является сложной проблемой в современной медицине. У ряда пациентов существующие методы лечения неэффективны, а трансплантация кишечника показывает неудовлетворительные результаты. Тканевая инженерия тонкой кишки может быть инновационным методом лечения этих пациентов. Одним из ключевых элементов в создании кишки является выращивание нервной системы кишечника. До настоящего времени не описаны культуры нервных клеток, выделенных из нервной системы кишечника, выращенные в трёхмерном матриксе на основе гиалуроновой кислоты. Цель исследования: разработать метод выращивания in vitro взаимодействующих друг с другом клеток нервной системы кишечника в трехмерной среде. Методы: клетки нервной системы кишечника выделялись из кишечника крыс путём препарирования и ферментативного воздействия на мышечный слой тонкой кишки. Далее клетки помещались в трёхмерный матрикс и культивировались в нём в течении нескольких дней. Результаты. Клетки нервной системы кишечника были выделены из крыс и культивировались в трёхмерном матриксе, помещённом в раствор для культур клеток с добавлением специфических факторов роста. Микроскопирование культур клеток на восьмые сутки показало, что нервные клетки активно растут, соединяясь с другими нервными клетками при помощи отростков. Конфокальная микроскопия в сочетании с иммуногистохимическим окрашиванием клеток специфичным маркером нервных клеток показала, что полученные культуры действительно состоят из нервных клеток. Заключение. Разработанный метод позволяет вырастить in vitro живущие и взаимодействующие друг с другом клетки нервной системы кишечника в трехмерной среде. Полученная культура клеток может быть использована для моделирования стенки тонкой кишки путем тканевой инженерии. Background. Treatment of short bowel syndrome is a complex issue in modern medicine. Existing treatment methods are inefficient in some cases, and bowel transplantation shows unsatisfactory results. Tissue engineering of the small intestine can represent an innovative method of treatment for these patients. One of the key elements in creation of the gut is culturing of enteric nervous system cells. There is no description of the nerve cells culture extracted from the enteric nervous system and grown in a hyaluronic-acid based three-dimensional matrix so far. Research objective: establishing an in vitro method for culturing of interacting enteric nervous system cells in a 3D environment. Methods. The enteric nervous system cells used for these experiments were isolated from Sprague-Dawley rats. Hyaluronic-acid based hydrogel HyStem®-C (ESI BIO - A Division of BioTime, USA) was used as a 3D matrix. Enteric nervous system cells were isolated from 5- to 8-day-old newborn Sprague-Dawley rats. Animals were decapitated with a rodent guillotine (cervical spinal cord transection). After that they were laparotomized, and their intestines were isolated. The removed intestines were placed in a Petri dish filled with the MEM (Minimum Essential Medium) supplemented with antibiotics (25 µl factory-produced solution of gentamycin (40 mg/ml) and 50 µl of metronidazole solution (5 mg/ml) per 50 ml of MEM). The small intestine was then separated from the colon and mesentery under an optical microscope, and the muscular layer of small intestine was isolated from the submucosal layer. The isolated muscular layer was placed in a tube with the deoxyribonuclease and collagenase solution as well as Balanced Salt Solution and incubated for 2 h at 37°C and 5% CO2. As a result, the intestinal muscle tissue was broken down completely in the produced solution, while areas of the myenteric plexus remained intact and were recognizable under the microscope as a network. These neural networks were treated with a trypsin solution. Further, after mechanical processing, a suspension of nerve cells was obtained. The hyaluronic-acid based gel HyStem®-C (ESI BIO - A Division of BioTime, Inc., USA) with added collagen was used to produce a 3D matrix. The enteric nervous system cells were added to this three-dimensional matrix and stirred. Matrix had been hardening for 30 minutes. The cell cultures were fixed, stained and microscoped 10-21 days later. For immunofluorescence staining of cells we used a direct immunohistochemical method with the anti-ß III Tubulin antibody conjugated with the fluorochrome Alexa 488 Flour (Merck Millipore). Anti-ß III Tubulin is a specific antibody used for peculiar staining of neurons. The anthraquinone dye with a high affinity to double-stranded DNA - DRAQ5 (Thermo Fisher Scientific) was applied in order to identify cell nuclei. Microscopy was conducted with Leica TCS SP8 (Leica, Germany). Results. The technique we represented allowed us to produce a culture of enteric nervous system cells isolated from rats in the three-dimensional matrix. Confocal microscopy combined with immunohistochemical staining with specific neuronal marker showed that the final cultures indeed consisted of nerve cells. Larger nerve cell clusters were encountered. Herewith, some neurons were arranged close to each other while others were located at a distance; however, all neurons were interconnected. Neurons also clearly showed wide processes morphologically similar to those of axons. In addition, many thin and branching processes resembling dendrites morphologically were visible. The use of confocal microscopy allowed to receive a series of images at different depths of the focal plane, which could then be reconstructed to a three-dimensional image. The 3D reconstruction of the nervous plexi clearly revealed neurons joined together in a complex network. For DRAQ5 stains all cell nuclei, not just those of neurons, the presence of stained cell nuclei just “hanging” on the reconstruction without cytosomes of nerve cells indicated that other cell types were also present in this texture. The present work showed a possibility for culturing interconnected enteric nervous system cells as well as nerve plexi within the three-dimensional medium in vitro. The key step in growing the enteric nervous system cells for both two-dimensional and three-dimensional medium claims to be the characteristics of functional cellular activity. Following stages of the study require observing of the cells’ functionality, for example, by using the analysis of intracellular Ca 2+ concentration (calcium imaging) and its variability. In addition, the study of cellular interactions within the enteric nervous system with other types of cells in the three-dimensional medium also encourages a great interest. Conclusion. The developed method enables one to culture the interconnected cells of the enteric nervous system within the three-dimensional medium in vitro, which can be used to create the enteric nervous system when growing the small intestinal wall by means of tissue engineering. Further research is based on studying the functionality of cells grown in a three-dimensional medium as well as on co-culturing of enteric nerve cells with other types of cells in three-dimensional media.
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Basu, Anustup. "The Passion of the Digital: the Ontology of the Photographic Image in the Age of New Media." Recherches sémiotiques 31, no. 1-2-3 (November 20, 2014): 175–202. http://dx.doi.org/10.7202/1027447ar.
Повний текст джерелаАнотація:
Using Mel Gibson’s 2004 film and cultural phenomenonThe Passion of the Christas a launching pad, this essay meditates on some questions about the twentieth century legacy of competing realisms, the graphic imperative of contemporary digital image cultures, and the ontological conundrums involving technology and mass media.Passionis an onto-theological filmic ‘event’ that derives equally from an almighty religiosity as well as a cultish process of being enraptured by certain ritual values of a new-age technologism of sound and image. This endographic writing out of the Gospel narrative at the level of the tissue and nerve of the committed viewer affirms a transcendental truth already there in an internal cosmos of belief instead of working in terms of an externally navigable ‘realist’ representation of the world that seeks to ‘bear away our faith’. This is rendered possible when unquestioning belief in Christ and in his momentous sacrifice is met by an embracing of technologywithout the skepticism of a scientific temper.
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Seil, Fredrick J. "Tissue Culture Models of Myelination After Oligodendrocyte Transplantation." Journal of Neural Transplantation 1, no. 2 (1989): 49–55. http://dx.doi.org/10.1155/np.1989.49.
Повний текст джерелаАнотація:
Studies of myelination after transplantation of mature oligodendrocytes to cerebellar cultures in which oligodendrocyte maturation and myelination had been irreversibly inhibited by exposure to cytosine arabinoside were reviewed. Transplanted oligodendrocytes were derived from three sources, including cerebellar explants treated with kainic acid, dissociated oligodendrocyte cultures, and optic nerve fragments. Oligodendrocytes from all sources migrated into the host explants and myelinated appropriate axons. The time of appearance of myelin and the percentage of host cultures myelinated differed for the three sources of oligodendrocytes, however. Myelin was visible earliest and in the highest percentage of host explants transplanted with cultured dissociated oligodendrocytes, which were presumably the most free to migrate into the host tissue, and latest and in the lowest percentage of host cultures transplanted with optic nerve, from which oligodendrocytes were presumably least free to migrate. Some myelin-like membranes unassociated with axons appeared in cerebellar cultures transplanted with cultured dissociated oligodendrocytes, and not in cerebellar explants transplanted with oligodendrocytes from other sources. The formation of such myelin-like membranes was interpreted as a manifestation of oligodendrocyte hyperreactivity induced by culture in isolation.
Дисертації з теми "Nerve tissue Cultures and culture media"
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McFadden, Thomas Bernard. "Influence of hormones on synthesis and secretion of milk proteins by mammary tissue from male and female cattle of beef and dairy breeds." Thesis, Virginia Polytechnic Institute and State University, 1985. http://hdl.handle.net/10919/101238.
Повний текст джерелаАнотація:
The ability of mammary tissue from prepubertal bulls and heifers of beef and dairy breeds to respond to hormonal stimuli through synthesis and secretion of milk proteins was studied. Experimental animals were six to eight month old Angus and Holstein cattle. All subjects were injected with estradiol and progesterone for seven days and slaughtered on day 15. Mammary tissue was explanted and cultured for 96 h in basal medium (B) which contained hormones necessary for maintenance, or stimulatory medium (P), further supplemented with prolactin. Selected cultures were incubated for 24 h in B or P medium containing 3H-amino acids. Concentrations of non-labeled alpha-lactalbumin (Alac), 3H-Alac, and 3H-total protein (TP) were determined in media and in explant homogenates.
Among cultures of bull mammary tissue, Angus explants secreted greater overall quantities of 3 H-TP and 3H-Alac than Holstein explants (p<.05). Secretion of Alac was also greater in Angus cultures at two of eight treatment periods (p<.01). Concentrations of all three protein fractions were likewise enhanced in homogenates of Angus explants for at least three of four treatment periods (p≤.05). Presence of prolactin in medium stimulated secretion of Alac (p<.005), and accumulation of all three fractions in explants (p<.10).
Holstein heifer explants secreted more Alac at three of eight treatment periods than Angus explants (p<.0005). Overall secretion of ³H-TP and ³H-Alac also was elevated in Holstein over Angus females (p<.10), as were concentrations of all three fractions in homogenates (p≤.01). Presence of prolactin had no direct effect on any protein parameters in female tissue.
I conclude that mammary tissue of immature bulls and heifers can be hormonally induced to express it's genetic merit for milk production (based on breed differences), through synthesis and secretion of milk proteins. Prolactin stimulated protein production in bulls but not in heifers. These findings indicate that similar methods of stimulating mammary tissue to produce milk proteins may be adaptable for commercial evaluation of genetic potential for milk production, especially in young bulls.M.S.
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Vernekar, Varadraj Nagesh. "Optimization of 3-d neural culture and extracellular electrophysiology for studying injury-induced morphological and functional changes." Diss., Georgia Institute of Technology, 2010. http://hdl.handle.net/1853/39624.
Повний текст джерелаАнотація:
This work characterized an in vitro 3-D neural co-culture model electrophysiologically via multi electrode arrays (MEAs), and morphologically via immunocytochemistry. Since MEA surface insulation SU-8 2000 can be used in neural micro- and multi- electrode arrays, this investigation first developed techniques to make SU-8 2000 cytocompatible. The in vitro 3-D neural co-culture model was then used to study viability and electrophysiological responses to physical injury as well as drugs known to affect network signaling. 1) SU-8 2000 cytotoxicity to neuronal cultures was linked to both poor adhesive properties and toxic components, such as solvents and photo acid generator elements. Surface treatments of oxygen plasma or parylene coating following optimal combinations of heat and isopropanol sonication showed improvement in SU-8 2000 cytocompatibility. 2) The 3-D neural networks within the 3-D co-cultures maintained considerable process outgrowth and complex 3-D structure. The cultures were viable up to three weeks in vitro with functional synaptic connections and spontaneous electrophysiological activity that was responsive to chemical modulation. This electrophysiological activity was modulated by synaptic inhibition. 3) Injury experiments demonstrated that both shear and compression loading significantly increased acute membrane permeability of cells in a strain rate dependent manner. Cell death correlated with higher membrane permeability, and shear resulted in more death than compression in these 3-D cultures. While techniques were developed for making a major micro-fabrication material cytocompatible, engineering the 3-D neural co-culture resulted in a more physiologically-representative neural tissue platform, allowing an increased understanding of structure-function relationships. Overall, this research established and characterized a neural culture system for the mechanistic study of cell growth, cell-cell and cell-matrix interactions, as well as the responses to chemical or mechanical perturbations. This is the first investigation of the network-level electrophysiological activity of 3-D dissociated cultures. This system can be used to model various pathological states in vitro, testing various reparative drugs; cell-, and tissue-engineering based strategies; as well as for pre-animal and pre-clinical testing of neural implants.
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Dinis, Tony Mickael. "Prothèse nerveuse artificielle à partir de fibroïne de soie pour la réparation et la régénération de nerfs périphériques." Thesis, Compiègne, 2014. http://www.theses.fr/2014COMP2152/document.
Повний текст джерелаАнотація:
La lésion de nerfs périphériques peut engendrer des déficits moteurs et/ou sensoriels permanents. En dépit des progrès techniques réalisés au cours de ces 25 dernières années, une récupération complète suite à ces lésions n’est pas encore possible aujourd'hui. L’autogreffe nerveuse, toujours considérée comme le standard clinique, est la seule technique capable d’offrir les meilleurs résultats en termes de récupération fonctionnelle. Cependant, la survenue de complications post-opératoires lors d’autogreffes d’un nerf et la quantité limitée de nerfs disponibles conduisent à mettre au point d’autres stratégies alternatives. Dans ce contexte, la mise au point de biomatériaux pour substituts nerveux devient une nécessité clinique. Malgré les efforts de la recherche, ces prothèses ne permettent toujours pas une régénération du nerf à la hauteur de l’autogreffe. Le biomatériau utilisé doit notamment présenter des propriétés physiques et chimiques proches de celui du nerf natif. La soie, aux propriétés mécaniques uniques, représente une bonne alternative pour mettre au point ce type de prothèses. En effet, la protéine de soie déjà utilisée dans le domaine biomédical est biocompatible. Les modifications chimiques de cette protéine améliore et favorise l’adhérence et la croissance cellulaires par l’incorporation de facteurs de croissance ou d’autres molécules d'intérêt. Ce travail de thèse propose de développer un nouveau type de biomatériau à base de soie fonctionnalisée par deux facteurs de croissance : le Nerve Growth Factor (NGF) et le Ciliary NeuroTrophic Factor (CNTF). Étant donné l’architecture complexe qui compose la structure nerveuse, une matrice supportant la repousse des tissus de façon orientée semble primordiale. Nous démontrons, dans un premier temps, le pouvoir de ces nanofibres alignées (produites par electrospinning) à orienter la régénération tissulaire de différents organes par culture d’explants. Les nanofibres de soie alignées, biocompatibles sont bio-activées par ajout de NGF spécifique de la régénération nerveuse. Cette matrice créée présente un gradient de concentration en NGF qui permet d’orienter la repousse axonale en stimulant la croissance axonale dans une seule direction. Afin d’optimiser la croissance de deux populations cellulaires, nous avons incorporé du CNTF pour produire des nanofibres bifonctionnalisées. Ces nanofibres bifonctionnalisées ont conduit à une longueur des neurites 3 fois plus grande à leurs contacts, stimulant la croissance des cellules gliales. Ainsi, nous avons produit des conduits nerveux à base de soie biofonctionnalisée pour implantation chez le rat. Les analyses physico-chimiques et les propriétés mécaniques démontrent le caractère biomimétique de nos tubes de guidage. Les premières études de la locomotion et l’observation de coupes du nerf sciatique de rat, suite à l’implantation de nos conduits donnent des résultats très prometteurs. L’ensemble de ces travaux démontre l’efficacité de nos guides nerveux à base de soie et les présente comme une alternative prometteuse à l’autogreffe nerveuse pratiquée en cliniquePeripheral nerve injury causes sensory and/or motor functions deficits. Despite technological advances over the past 25 years, a complete recovery from these injuries remains unsatisfactory today. The autograft still considered the "gold standard" in clinical practice. This is the only technique able to offer complete functional recovery. However, the occurrence of postoperative complications in autologous nerve and the limited amount of available nerves lead to develop alternatives strategy.In this context, development of nerve graft substitutes becomes by far a clinical necessity. Despite research efforts, these artificial prostheses design based on biomaterial doesn’t allow nerve regeneration as found in autograft nerve procedures. The biomaterial used must have the physical and chemical properties similar to that of the native nerve. Silk, well known for its unique mechanical properties, proposes a good alternative to develop these prostheses. Indeed, the silk protein is commonly used in the biomedical field and regenerative medicine. This protein biocompatibility may be improved through chemical modifications to promote adhesion and cell growth by the incorporation of growth factors or other molecules of interest. Therefore, this thesis proposes to develop a new type of functionalized silk biomaterial based on two growth factors : Nerve Growth Factor (NGF) and Ciliary NeuroTrophic Factor (CNTF). Given the complex architecture that consists of nerve structure, a matrix which is able to support and manage the outgrowth of tissue becomes essential. We demonstrate the power of these aligned nanofibers (produced by electrospinning) to guide and manage tissue regeneration from different organ explants culture. Aligned silk nanofibers, were biocompatible and bio-activated by adding NGF involved for nerve regeneration. This matrix has been created with a concentration gradient of NGF to guide neuritis outgrowth in only one direction. The presence of this gradient demonstrated a better axonal growth in one direction versus the uniform concentration conditions. Nerve cells consist essentially of two cell populations which are neurons and Schwann cells. To optimize the culture and growth of these two populations, in addition to NGF, we incorporated CNTF to produce bifunctionalized nanofibers. These biofunctionalised nanofibers led to a length 3 times larger on contact with neurites. The glial cells growth, alignment and migration were stimulated by CNTF. Thus, we produced bi-functionalized nerve guidance conduits for rat implantation. The physico-chemical analyzes demonstrate the biomimetic of our guide tubes. Early studies of locomotion and observing histological sections of rat sciatic nerve, following the implementation of our conduits gave very promising results.These studies demonstrate the relevance of our nervous guides’ silk-based developed as an effective alternative to nerve autograft performed in the clinic
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Zhi, Chunxing. "Cultivation of Laminaria saccharina gametophyte cell cultures and Acrosiphonia coalita tissue cultures in a bubble-column photobioreactor." Thesis, 1994. http://hdl.handle.net/1957/35536.
Повний текст джерелаКниги з теми "Nerve tissue Cultures and culture media"
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W, Haynes L., ed. The neuron in tissue culture. Chichester: Wiley, 1999.
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Abraham, Shahar, ed. A Dissection and tissue culture manual of the nervous system. New York: Liss, 1989.
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Protocols for neural cell culture. 4th ed. New York: Humana Press, 2010.
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Sergey, Fedoroff, and Richardson Arleen, eds. Protocols for neural cell culture. 3rd ed. Totowa, N.J: Humana Press, 2001.
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Mitsuhashi, Jun. Invertebrate tissue culture methods. Tokyo: Springer, 2002.
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Mitsuhashi, Jun. Invertebrate tissue culture methods. Tokyo: Springer, 2002.
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International, Conference on Invertebrate and Fish Tissue Culture (8th 1991 Anaheim Calif ). Eighth International Conference on Invertebrate and Fish Tissue Culture: Proceedings of the Eighth International Conference on Invertebrate and Fish Tissue Culture, held in conjunction with the 1991 World Congress on Cell and Tissue Culture, June 16-20, 1991, Anaheim, California. Columbia, Md., USA: Tissue Culture Association, 1992.
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International Conference on Invertebrate and Fish Tissue Culture (7th 1987 Ohito, Japan). Invertebrate and fish tissue culture: Proceedings of the Seventh International Conference on Invertebrate and Fish Tissue Culture, Japan, 1987. Tokyo: Japan Scientific Societies Press, 1988.
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Davies, Jamie A. Replacing animal models: A practical guide to creating and using culture-based biomimetic alternatives. Chichester, West Sussex: John Wiley & Sons, 2012.
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A, Boulton A., Baker Glen B. 1947-, and Walz Wolfgang, eds. Practical cell culture techniques. Totowa, N.J: Humana Press, 1992.
Знайти повний текст джерелаЧастини книг з теми "Nerve tissue Cultures and culture media"
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Mitsuhashi, J. "Simplification of Media and Utilization of Sugars by Insect Cells in Cultures." In Invertebrate and Fish Tissue Culture, 15–18. Berlin, Heidelberg: Springer Berlin Heidelberg, 1988. http://dx.doi.org/10.1007/978-3-642-73626-1_4.
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Gstraunthaler, G., T. Seppi, C. Monteil, E. Healy, M. P. Ryan, J. P. Morin, and W. Pfaller. "The impact of growth conditions, culture media volume and glucose content on differentiation and metabolism of renal epithelial tissue cultures." In Ersatz- und Ergänzungsmethoden zu Tierversuchen, 265–66. Vienna: Springer Vienna, 1998. http://dx.doi.org/10.1007/978-3-7091-7500-2_45.
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Mukherjee, Nabanita, Karoline A. Lambert, David A. Norris, and Yiqun G. Shellman. "Enrichment of Melanoma Stem-Like Cells via Sphere Assays." In Methods in Molecular Biology, 185–99. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1205-7_14.
Повний текст джерелаАнотація:
AbstractSphere assays are widely used in vitro techniques to enrich and evaluate the stem-like cell behavior of both normal and cancer cells. Utilizing three-dimensional in vitro sphere culture conditions provide a better representation of tumor growth in vivo than the more common monolayer cultures. We describe how to perform primary and secondary sphere assays, used for the enrichment and self-renewability studies of melanoma/melanocyte stem-like cells. Spheres are generated by growing melanoma cells at low density in nonadherent conditions with stem cell media. We provide protocols for preparing inexpensive and versatile polyHEMA-coated plates, setting up primary and secondary sphere assays in almost any tissue culture format and quantification methods using standard inverted microscopy. Our protocol is easily adaptable to laboratories with basic cell culture capabilities, without the need for expensive fluidic instruments.
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Gould, Jean. "[30] Nondestructive assay for β-glucuronidase in culture media of plant tissue cultures." In Methods in Enzymology, 357–62. Elsevier, 1992. http://dx.doi.org/10.1016/0076-6879(92)16032-f.
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Wrzesinski, Krzysztof, Søren Alnøe, Hans H. Jochumsen, Karoline Mikkelsen, Torsten D. Bryld, Julie S. Vistisen, Peter Willems Alnøe, and Stephen J. Fey. "A Purpose-Built System for Culturing Cells as In Vivo Mimetic 3D Structures." In BioMechanics and Functional Tissue Engineering [Working Title]. IntechOpen, 2021. http://dx.doi.org/10.5772/intechopen.96091.
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Culturing cells in 3D is often considered to be significantly more difficult than culturing them in 2D. In practice, this is not the case: the situation is that equipment needed for 3D cell culture has not been optimised as much as equipment for 2D. Here we present a few key features which must be considered when designing 3D cell culture equipment. These include diffusion gradients, shear stress and time. Diffusion gradients are unavoidably introduced when cells are cultured as clusters. Perhaps the most important consequence of this is that the resulting hypoxia is a major driving force in the metabolic reprogramming. Most cells in tissues do not experience liquid shear stress and it should therefore be minimised. Time is the factor that is most often overlooked. Cells, irrespective of their origin, are damaged when cultures are initiated: they need time to recover. All of these features can be readily combined into a clinostat incubator and bioreactor. Surprisingly, growing cells in a clinostat system do not require specialised media, scaffolds, ECM substitutes or growth factors. This considerably facilitates the transition to 3D. Most importantly, cells growing this way mirror cells growing in vivo and are thus valuable for biomedical research.
Тези доповідей конференцій з теми "Nerve tissue Cultures and culture media"
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Mignone, Lindsay F., Shirley Masand, Jeffrey D. Zahn, and David I. Shreiber. "A Simple, Cost-Effective Method to Improve Cell Viability in Microniche Culture Systems." In ASME 2010 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2010. http://dx.doi.org/10.1115/sbc2010-19189.
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Microfluidic networks are increasingly used to generate custom microenvironmental niches for cell culture and assays of cellular behavior. Perfusion systems are typically required to overcome diffusive limitations associated with culturing cells longer than a few hours when nutrient delivery, oxygen delivery and metabolic waste removal are required to maintain cell viability. In addition to the added complexity of experimental methods, perfusion systems can result in nonuniform nutrient delivery and subject cells to shear stresses, which may alter cell behavior and possibly cause cell death. In particular, when culturing cells within hydrogel scaffold-filled networks, as may be done in micro-tissue engineering, the need for perfusion culture also increases the likelihood of a destructive bubble entering the network. Moreover, analysis of micro-cultures frequently entails labelling with antibodies and/or fluorescent probes, which again requires controlled perfusion of the various reagents through the network. We have developed a simple technique to preserve cell viability and simplify labeling within microscale cultures without the need for perfusion. Instead of bonding a microfluidic network to glass, PDMS, or another impermeable substrate, the network is bonded to a semi-permeable microdialysis membrane, which allows free exchange of oxygen, proteins, nutrients, and waste between the microfluidic channels and culture media in static culture plates.
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Lelkes, Peter I., and Brian R. Unsworth. "Cellular Signaling Mechanisms Involved in the 3-Dimensional Assembly and Differentiation of PC12 Pheochromocytoma Cells Under Simulated Microgravity in NASA Rotating Wall Vessel Bioreactors." In ASME 1998 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 1998. http://dx.doi.org/10.1115/imece1998-0791.
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Abstract Rotating Wall Vessel (RWV) Bioreactors simulate microgravity and facilitate 3-D tissue-like assembly through spatial co-localization and cell-cell interactions. This unique cell culture venue is well suited to assess the role of micro-environmental cues in the assembly and tissue-specific differentiation of cells in culture. Our long term goal is to use RWV Bioreactors for generating functional neuroendocrine 3-D constructs which may be useful as clinical replacement tissue in treating neurodegenerative diseases. As a model we are using PC12 pheochromocytoma cells, a bipotential rat adrenal medullary tumor cell line. PC 12 cells differentiate, depending on exogenous factors, either along the neuronal or the neuroendocrine pathway. PC12 cells, when maintained for up to 20 days in RWV Bioreactors, form macroscopic tissue-like aggregates which exhibit enhanced expression of neuroendocrine, adrenergic differentiation markers (Lelkes et al., In Vitro Devel. Biol, 1998, 34: 316–325). We hypothesized that exposure of PC12 cells to the “simulated microgravity” culture conditions in the RWV Bioreactors, might selectively activate signal transduction pathways leading to enhanced neuroendocrine adrenergic differentiation. Using quantitative RT-PCR we demonstrated rapid upregulation of an adrenergic marker, phenylethanolamine-N-methyl transferase (PNMT), in short term RWV cultures. Concomitantly, we found, by electrophoretic mobility shift assays, differential induction of nuclear transcription factors, such as GRE and SP-1, which are known to be involved in the glucocorticoid-induced activation of PNMT. Conversely, upon short term culture of PC12 cells in RWV, the neuronal traits of the cells were impaired. Upon exposure to simulated microgravity, MAPK signaling (erk and jnk) was constitutively activated, while nerve-growth factor (NGF)-induced activation of erk, was abrogated. These results suggest that the culture conditions in the RWV Bioreactors are sufficient to induce PC12 cell differentiation towards the neuroendocrine, phenotype by upregulating “adrenergic” gene expression, while downregulating neurotrophin signaling pathways.
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Merryman, W. David, Kristen Loveless, and Mehran Kasra. "Disc Nucleus Cellular Response to Dynamic Pressures at Critical Frequencies: A Pig Model." In ASME 2003 International Mechanical Engineering Congress and Exposition. ASMEDC, 2003. http://dx.doi.org/10.1115/imece2003-43092.
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Disc degeneration is a multifactor phenomenon. It has been found that intervertebral disc (IVD) cells respond to such factors as pH, osmotic pressure, genetic factors, and mechanical loading (Guilak, 1999). During daily activities the human intervertebral disc is exposed to oscillatory hydrostatic loads that produce pressures >2MPa in vivo (Nachemson, 1964 and 1979). It is known that dynamic loads with critical frequencies close to that of the in vivo human spine resonant frequency (4–5 Hz) have a destructive effect on disc tissue properties (Pope, 1993). Whether this destructive effect is purely mechanical, due to loading magnification, or biological, affecting cell metabolism, is unknown. Previous work (Merryman, 2002) showed that there was no significant effect upon monolayer IVD cells loaded at 15Hz, while lower frequencies (1 and 8Hz) altered collagen synthesis compared to control. To address this issue, we developed a mechanically active culture system capable of delivering a wide range of loading frequencies and amplitudes of hydrostatic pressure to cultures of disc cells. Nucleus pulposus cells of pig discs were isolated and suspended in alginate beads. Alginate cultures were divided into 6 groups; five groups were exposed to cyclic pressures of frequencies 1, 3, 5, 8, and 10Hz with the same amplitude of 1MPa, and group 6 was the control group (no loading). Cultures of different groups were loaded for 3 days (30 minutes daily) in a hydraulic chamber filled with culture media. The effect of loading frequency on collagen metabolism among different groups was compared by measuring incorporated [3H]-proline into collagen for medium and total extracts. The results indicated a poor synthesis rate and more degradation near the 5Hz frequency.
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Xia, Chunguang, and Nicholas Fang. "Enhanced Mass Transport Through Permeable Polymer Microcirculatory Networks." In ASME 2006 International Mechanical Engineering Congress and Exposition. ASMEDC, 2006. http://dx.doi.org/10.1115/imece2006-15408.
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One of the obstacles of culturing functioning vital tissues in vitro is to obtain a substantial biomass at a physiological cell density (>108cells/cm3). At this high density, the diffusion length of metabolites is limited to ~100um. As a matter of fact, in real tissue, almost all the cells are located within 100um distance from the capillaries [1]. Studies [2, 3] also confirmed that the cells in the artificial tissue cannot be properly cultured when they are further than 400um from the external nutrient source. Therefore, to culture three dimensional artificial tissue with substantial biomass, vascularization is necessary to enhance the metabolites transport. The short diffusion length of the metabolites requires high capillary density (>100/mm2) in vascularization. To meet this need, we have developed a novel high resolution and high speed 3D microfabrication technique, the projection microstereolithography[4] to explore microcirculatory networks with high density (>150/mm2). Using this technology we designed and fabricated the microreactors as shown in Figure 1. In our samples, 800um PEG microcapillaries with 20um inner radius and 40um outer radius with pitch of 96um are fabricated. Two rings as inlet and outlet are connected to external supply of culture medium. We designed the parameters of the vascularized microbioreactor based on the simulations of oxygen and carbon dioxide transport and metabolism in hepatocytes. As shown in Figure 2, the capillaries are arranged in a hexagonal way. According to the geometric symmetry, the final simulation domain is divided into 2 regions, the polymer capillary wall and the tissue. We assumed that a culture media with dissolved oxygen is pumped through the capillaries at 1.5mm/s rate and diffuses through the capillary wall, into the hepatocytes. The consumption of oxygen follows Michaelis-Menten kinetics [5, 6] and the metabolic rate of carbon dioxide is assumed to be proportional to that of oxygen by a fixed quotient (-0.81) which is addressed and studied by other groups [7]. The carbon dioxide diffuses into the capillaries and can be carried away through the flow of the culture medium. Our simulation indicates that the bottleneck of effective oxygen transport is the permeability of the polymer materials. The oxygen concentration drops off about 90% after diffusing through the capillary wall. It is predicted that the diffusion length at the inlet is 74um and 48um at the outlet; the rapid drop of carbon dioxide concentration also happens across the capillary wall. The predicted carbon dioxide concentration in the tissue is ~80nmol/cm3; this value is much smaller than the toxic value (100mmHg or 3umol/cm3) reported by David Gray and coworkers [8]. In Figure 2, we present the effect of the permeability of the capillary polymer materials on the diffusion length of oxygen and the concentration of carbon dioxide in the tissue. Our study indicates the existence of an optimal permeability for the capillary network, at which the overall diffusion length of oxygen is maximized. Interestingly, we also found a maximum concentration of carbon dioxide in the cultured tissue as the permeability of the polymer material changes. We attribute it to the competition between the tissue thickness and the permeability. Higher permeability increases the cultured tissue thickness, and also increases the ability of capillary to empty carbon dioxide. Not only is this model applicable for oxygen and carbon dioxide, but also for the transport of other metabolites. As an ongoing experimental effort, our fluorescent microscopy measurement validated the diffusion transport of fluorescent species from the capillary (Figure 3). Experiments are also in progress on the oxygen diffusion from the capillaries will cell cultures of high density on the PEG scaffold by introducing proper indicators. In summary, we have established a method to design and manufacture vascularized microcirculatory network to enhance the mass transport during the tissue culture. To ensure the effective nutrient delivery and wastes removal, our numerical simulation also confirms that it is essential to embed high density microcapillaries with optimal permeability.
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Elbashir, Israa, Aisha Aisha Nasser J. M. Al-Saei, Paul Thornalley, and Naila Rabbani. "Evaluation of antiviral activity of Manuka honey against SARS-CoV-2." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2021. http://dx.doi.org/10.29117/quarfe.2021.0113.
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Background and aims: In 2020 a global pandemic was declared caused by the severe acute respiratory syndrome coronavirus (SARS-CoV-2). The pandemic is still ongoing and continues to cause considerable mortality and morbidity world-wide and new variants of the virus are emerging. Rapid development and rollout of vaccines for SARS-CoV-2 is in progress to counter the pandemic but has been tempered by the emergence of new SARS-CoV-2 variants, many of which exhibit reduced vaccine effectiveness. To date there is no approved antiviral treatment for coronavirus disease 2019 (COVID-19). Several studies have shown that Manuka honey has virucidal/antiviral effect. Methylglyoxal (MG), a bioactive component in Manuka honey, has antiviral activity in vitro. MG may modify arginine residues in the functional domains of viral spike and nucleocapsid proteins, resulting in loss of charge, protein misfolding and inactivation. The aim of this study was to characterize the antiviral activity of Manuka honey against SARS-CoV-2 in vitro Materials and methods: Wild-type SARS-CoV-2 with titers of multiplicities of infection (MOI) 0.1 and 0.05 were incubated with 2-fold serial dilutions of 250+ Manuka honey (equivalent to 250 to 31 µM) in infection medium (Dulbecco's Modified Eagle Medium + 2% fetal bovine serum + 100 units/ml penicillin + 100 µg/ml streptomycin) for 3 h. Manuka honey treated and untreated control SARS-CoV-2 was incubated with confluent cultures of Vero cells in vitro for 1 h, cultures washed with phosphate-buffered saline and incubated in fresh infection medium at 37°C for 4 - 5 days until 70% of virus control cells displayed cytopathic effect. We also studied the effect of scavenging MG in Manuka Honey with aminoguanidine (AG; 500 µM) on virucidal activity. The antiviral activity of MG was judged by median tissue culture infectious dose (TCID50) assays. Data analysis was by logistic regression. TCID50 (mean ± SD) was deduced by interpolation. Results: Diluted Manuka honey inhibited SARS-CoV-2 replication in Vero cells. SARS-CoV-2 was incubated in diluted Manuka honey in medium at 37°C for 3 h before adding to Vero cells. Manuka honey dilutions down to 125 µM MG equivalents completely inhibited cytopathic effect of SARS-CoV-2 whereas 31.25 µM and 62.5 µM MG equivalents had limited effect. Logistic regression and interpolation of the cytopathic effect indicated that the TCID50 = 72 ± 2 µM MG equivalents for MOI of 0.1. Prior scavenging of MG by addition of AG resulted in virus replication levels equivalent to those seen in the virus control without AG. Conclusion: Manuka honey has antiviral activity against SARS-CoV-2 when incubated with the virus in cell-free media at no greater than ca. 40-fold dilutions of 250+ grade. Anti-viral activity was inhibited by AG, consistent with the anti-viral effect being mediated by MG. Manuka honey dilutions in MG equivalents had similar antiviral effect compared to authentic MG, also consistent with MG content of Manuka honey mediating the antiviral effect. Whilst Manuka honey may inactivate SARS-CoV-2 in cell-free culture medium, its antiviral activity in vivo for other than topical application may be limited because of the rapid metabolism of MG by the glyoxalase system and limited bioavailability of oral MG.