REVIEW pre Culture conditions for hESC

Thinking of the quality of the cells since there is

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Unformatted text preview: quality of the cells since there is no contact of blastocyst with animal-derived pronase, antibodies, and complement factors. The latter procedure has been recently adapted for the establishment of new hESC line by Genbacev et al. (2005). We have also derived our latest five hESC lines mechanically (Fig. 1). Feeder cells in human embryonic stem cell cultures The establishment of hESC lines was first described by using mouse embryonic fibroblasts (MEFs) as feeder cells and fetal bovine serum (FBS)-containing culture medium for both feeder cells and hESC (Thomson et al. 1998). For replacing animal cells, various alternative human cell lines have been successfully tested as feeder cells for hESC. Table 2 summarizes the sources of human feeders that have been validated for culture of undifferentiated hESC. Richards and co-workers have studied different kinds of human cells, such as human fetal muscle and skin, human adult Fallopian tube epithelial feeders, and adult muscle cells as feeders for hESC (Richards et al. 2002, 2003). A finding from their results was that for some reason MEF cells seemed to support hESC growth better than human feeders. Our group has derived and cultured all our 25 h ESC lines using commercially available human foreskin fibroblasts (hFF) as feeder cells (Hovatta et al. 2003, Inzunza et al. 2005). In our cultures, hFF seem to support the derivation and undifferentiated growth of hESC well. On the other hand, we have not compared the growth rates of hESC cultured with hFF or MEFs as feeder cells. The commercially available hFF lines (and other reported lines) have been derived and cultured in FBS-containing medium which often has batch-to-batch variation causing variation to culture conditions. Also, human adult marrow stroma cells for hESC maintenance (Cheng et al. 2003), and uterine endometrium cells (Lee et al. 2005) as well as human placenta fibroblasts (Genbacev et al. 2005) for derivation and maintenance of hESC have been used, but all these feeder cells have been cultured using FBS in culture medium. Table 2 Human cells used as feeder cells for human embryonic stem cell cultures. Feeder cell source Human foreskin Placenta Uterine endometrium Adult marrow stroma hESC-derived fibroblasts Fetal muscle Fetal skin Adult fallopian tube Adult skin Adult muscle Feeder cell medium composition Derivation (D)/maintenance (M) FBS/HS FBS FBS FBS FBS FBS FBS M D/M D/M D/M D/M M M M D/M D/M D/M D/M M M HS HS Reference Amit et al. (2003) Hovatta et al. (2003) Inzunza et al. (2005) Genbacev et al. (2005) Lee et al. (2005) Cheng et al. (2003) Xu et al. (2004) Stojkovic et al. (2005b) Wang et al. (2005c) Richards et al. (2002) Richards et al. (2003) FBS, fetal bovine serum; HS, human serum; hESC, human embryonic stem cell. Reproduction (2006) 132 691–698 www.reproduction-online.org Culture conditions for hESCs 693 Table 3 Feeder-free culture methods established for human embryonic stem cells. Derivation (D)/ maintenance (M) Coating ma...
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This document was uploaded on 09/21/2013.

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