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REVIEW pre Culture conditions for hESC

D maintenance m coating material medium supplements

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Unformatted text preview: terial Medium supplements Matrigel/laminin Matrigel/laminin Matrigel Matrigel Matrigel Matrigel/laminin Matrigel/collagen IVC fibronectinClamininCvitronectin FBS coating Laminin Matrigel Matrigel Fibronectin Fibronectin ECM from MEFs HS matrix Laminin CM (MEFs), SR, bFGF CM (MEFs), SR, bFGF CM (hESCd-F), SR, bFGF SR, bFGF, GSK3 inhibitor CM (MEFs/HES), S1P, PDGF, bFGF X-VIVO 10, bFGF Defined, animal-free medium M M M M M M D/M Xu et al. (2001) Carpenter et al. (2004) Xu et al. (2004) Sato et al. (2004) Pebay et al. (2005) Li et al. (2005) Ludwig et al. (2006) Reference CDM, activin A, noudal, bFGF SR, activin A, KGF, NIC SR, bFGFCnoggin or high bFGF SR, bFGF, noggin, high bFGF SR, TGF-b, LIF, bFGF CM (MEFs), SR, bFGF SR, Plasmanate, bFGF CM (hESCd-F) X-VIVO10, high bFGF M M M M M M D/M M M Vallier et al. (2005) Beattie et al. (2005) Xu et al. (2005) Wang et al. (2005c) Amit et al. (2004) Noaksson et al. (2005) Klimanskaya et al. (2005) Stojkovic et al. (2005a) Genbacev et al. (2005) CM, conditioned medium; MEFs, mouse embryonic fibroblasts; SR, serum replacement; bFGF, basic fibroblast growth factor; hESC-dF, human embryonic stem cell derived fibroblasts; GSK3 inhibitor, glycogen synthase kinase-3 inhibitor; HES, human embryonic stem cell; S1P, sphingosine-1phosphate; PDGF, platelet derived growth factor; CDM, chemically defined medium; KGF, keratinocyte growth factor; NIC, nicotinamide; TGF-b, transforming growth factor b; LIF, leukaemia inhibitor factor; HS, human serum. The use of hESC-derived fibroblasts or fibroblast-like cells, and hESC-derived immortalized fibroblasts as a feeder cells has recently been described (Stojkovic et al. 2005b, Wang et al. 2005b). Xu et al. (2004) established a culture system where conditioned medium from hESCderived fibroblasts were used. Immortalized cell lines might provide feeder cells for extended time as compared with primary cell lines, such as adult fibroblast cell lines. Human ESC-derived feeders provide interesting opportunity where autogenic feeder cells could be derived from hESC and used for certain hESC lines. On the other hand, differentiation of fibroblast-like cells and using those for maintenance is very labor intensive and hardly the best way to standardize and produce mass cultures of undifferentiated hESC for research purposes. Systematic comparisons of the properties of hESC derived and grown on different feeders are needed in order to decide which feeder cells are optimal. In contrast, it may be possible that the hESC get adapted to the feeders on which they have been derived, which makes such comparison difficult. To avoid these problems, we and other researchers have optimized serum-free culture conditions for hESC lines using a basic fibroblast growth factors (bFGF) and a more defined serum replacement (KnockOut Serum Replacement, Invitrogen), which still contains animal proteins, which are not fully defined (see for example, Amit et al. 2000, Koivisto et al. 2004, Stojkovic...
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