REVIEW pre Culture conditions for hESC

REVIEW pre Culture conditions for hESC

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Unformatted text preview: tudies are missing. A number Conditioned medium from feeder cells or a medium with animal components (such as the widely used Invitrogen SR) is not optimal when we think of pathogens and possible immunoreactions as well as batch-to-batch variation of culture media. Very recently, important work has been carried out by replacing animal components in SR-containing medium with more defined media and supplements. Genbacev et al. (2005) and Li et al. (2005) used X-VIVO 10 with bFGF, Vallier et al. (2005) used chemically defined medium with activin A, nodal and bFGF, and Ludwig et al. (2006) used defined culture medium with bFGF, LiCl, GABA, TGF-b, and pipeolic acid. Defined quality cells also needed for research purposes Figure 2 Key features of human embryonic stem cell line culture conditions which need to be optimized. www.reproduction-online.org Reproduction (2006) 132 691–698 696 H Skottman and O Hovatta of karyotypic abnormalities have been identified in cultured hESC (Allegrucci & Young 2006). Human ESC is also epigenetically sensitive (Steele et al. 2005). In feeder-free culture of hESC, new gently passaging methods without dissociation of hESC for single cells might enable preventing abnormalities caused by selective pressure in long-term cultures of hESC. If the cells are cultured only for research purposes, we need not consider so much the immunoreactions caused by animal substances in culture medium, but pathogens may change the properties of the cells. The current culture methods are by no means perfect, and they need optimization also to gain high efficiency for the production of sufficient amounts of hESC for research purposes. The use of feeder cells sets limitations for research, since experimental data might result from a combination of hESC and feeder cells (Klimanskaya et al. 2005). The variation in culture protocols may also affect the ability to differentiate hESC with standard differentiation protocols or to provide standard systems for developmental studies, disease modeling, drug discovery, and toxicology applications. Recently established culture methods with high concentration of growth factors and use of human matrix components are very expensive for research purposes. Understanding the mechanisms of hESC self-renewal and proliferation as non-differentiated cells would greatly facilitate the establishment of defined derivation and culture conditions for hESC (Vallier et al. 2005, Wang et al. 2005a, Armstrong et al. 2006, Rho et al. 2006, Stewart et al. 2006). Recent advantages in the use of biomaterials (Anderson et al. 2004) and perfusion culture systems (Fong et al. 2005) for the culture of hESC suggest that large-scale culture of hESC in bioreactors under controlled environment using defined culture medium and biomaterial substrates may be feasible in future. International stem cell initiative (Andrews et al. 2005) has been established by several organizations which fund hESC research. It is an important atte...
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This document was uploaded on 09/21/2013.

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