Embryonic stem cells cell culture

Bain G, Ray WJ, Yao M et al (1996) Retinoic acid promotes neural and represses mesodermal gene expression in mouse embryonic stem cells in culture. Biochem Biophys Res Commun 223(3) 691-694... 

As long as embryonic stem cells in culture are grown under certain conditions, they can remain undifferentiated (unspecialized). But if cells are allowed to clump together to form embryoid bodies they begin to differentiate spontaneously into specific cell types (muscle cells, nerve cells, etc.). To generate cultures of specific types of differentiated cells, various growth factors are used in the culture media [45],... 

Thein-Han WW, Kitiyanant Y (2007) Chitosan scaffolds for in vitro buffalo embryonic stem-like cell culture an approach to tissue engineering. J Biomed Mater Res Appl Biomater 80B 92-101... 

Tada S, Era T, Furusawa C, Sakurai H, Nishikawa S, Kinoshita M, Nakao K, Chiba T. 2005. Characterization of mesendoderm a diverging point of the definitive endoderm and mesoderm in embryonic stem cell differentiation culture. Development 132(19) 4363 374. 

Nakano T, Kodama H, Honjo T. Generation of lymphohematopoietic cells from embryonic stem cells in culture. Science 1994 265 1098-101. 

By culturing specific cells from cloned embryos, scientists can make embryonic stem cell (ESC) cultures. During mammalian development, two distinct cell populations form after the first few days of embryonic development. The trophoblast, or the flattened, outer layer of cells, will eventually form the placenta and its associated structures. The inner cell mass (IGM) is the round, inner clump of cells that develop to form the embryo proper and a few structures associated with the placenta. If IGM cells are isolated and cultured on feeder cells, a layer of nondividing skin cells that secrete a cocktail of growth-promoting chemicals, the IGM cells will grow and spread over the surface of the culture dish. Such a culture is an embryonic stem cell culture, and these cells are pluripotent, which means that they can differentiate into any cell type in the adult body. 

Kubo, A., Shinozaki, K., Shannon, J. M. et al. 2004. Development of definitive endoderm from embryonic stem cells in culture. Development 131(7) 1651-62. 

T.M. Yoon, B. Chang, H.T. Kim, J.H. Jee, D.W. Kim, and D.Y Hwang, Human embryonic stem cells (hESCs) cultured under distinctive feeder-free culture conditions display global gene expression patterns similar to hESCs from feeder-dependent culture conditions. Stem Cell Rev. 6 (3) 425-437, Sep. 2010. 

Era, T., N. Izumi, M. Hayashi et al. 2008. Multiple mesoderm subsets give rise to endothelial cells, whereas hematopoietic cells are differentiated only from a restricted subset in embryonic stem cell differentiation culture. Stem Cells 26(2) 401-11. 

Dang, S. M., S. Gerecht-Nir, J. Chen, J. Itskovitz-Eldor, and P. W. Zandstra. 2004. Controlled, scalable embryonic stem cell differentiation culture. Stem Cells 22(3) 275-82. 

Matsui, Y Zsebo, K. M and Hogan, B. L. M. (1992). Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture. Cell 70 841-847. 

Culture of human embryonic stem cells starts with the recovery of the blastocyst s inner cell mass (Figure 14.17). One common recovery procedure is termed immunosurgery . The process... 

Figure 14.17 Overview of the generation and culture of human embryonic stem cells. IVF in vitro fertilization. Refer to text for further details...

Human embryonic stem cells / Culture as floating aggregratesN. (embryoid bodies), forming / neuroepithelial cells s Culture to form midbrain dopamine neurons ... 

Hoffman, L. and Carpenter, M. 2005. Characterization and culture of human embryonic stem cells. Nature Biotechnology 23(6), 699-708. 

Xu et al. [76] have showed that human embryonic stem cells by treatment with bone morphogenetic protein-4 can be driven to differentiate into tro-phoblasts which have the ability to syncytialize and form confluent mono-layers. The differentiated cells express a number of trophoblast markers and secrete placental hormones and thus may provide an alternative placental model. Under the culture conditions used, however, the cells propagated poorly. 

Zambidis E, Oberlin E, Tavian M and Peault B (2006). Blood forming endothelium in human ontogeny lessons from in utero development and embryonic stem cell culture. Trends in Cardiovascular Medicine 16(3) 95 101. 

Wartenberg M, Donmez F, Ling FC et al (2001) Tumor-induced angiogenesis studied in confrontation cultures of multicellular tumor spheroids and embryoid bodies grown from pluripotent embryonic stem cells. FASEB J 15 995-1005... 

Wartenberg M, Finkensieper A, Hescheler J et al (2006) Confrontation cultures of embryonic stem cells with multicellular tumor spheroids to study tumor-induced angiogenesis. Methods Mol Biol 331 313-328... 

In Europe, the developmental toxicity testing (including teratogenicity) of new cosmetic ingredients is performed according to the Cosmetics Directive 76/768/EEC only alternatives leading to full replacement of animal experiments should be used. This chapter presents the three scientifically validated animal alternative methods for the assessment of embryotoxicity the embryonic stem cell test (EST), the micromass (MM) assay, and the whole embryo culture (WEC) assay. 

Therefore, a validation exercise with a variety of compounds with unknown mechanisms of developmental toxicity has only limited value if only used to derive an overall predictability rate of a single assay. It is more useful to elucidate the applicability domain of the assay in terms of the mechanisms of development covered, and to validate that aspect by testing compounds that do or do not affect the applicability domain. For single end point assays such as specific receptor activation assays, this exercise is relatively straightforward. For more complex assays such as those involving embryonic cell differentiation, the understanding of the applicability domain is more complex, as extensive research with the embryonic stem cell test has learned (27, 47). Whole embryo cultures are probably more straightforward in terms of applicability domain as they involve the entire embryo in a limited window of development, but such assays are complex and not animal free. 

The embryonic stem cell test is an animal-free alternative test method for developmental toxicity. Mouse embryonic stem cells are cultured in a hanging drop method to form embryoid bodies. These embryoid bodies, when plated on tissue culture dishes, differentiate to form contracting myocardial cell foci within 10 days. Inhibition of cardiomyocyte differentiation by test compounds serves as the end point of the assay, as monitored by cormting contracting muscle foci under the microscope. 

---