REVIEW pre Culture conditions for hESC

REVIEW pre Culture conditions for hESC

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Unformatted text preview: or human feeder cells) (Xu et al. 2001, 2004). Matrigel is a complex mixture of mouse sarcoma origin which contains extracellular molecules, such as laminin, collagen IV, and also growth factors and other substances. Also other research groups have successfully used Matrigel as substrate for hESC (Sato et al. 2004, Pebay et al. 2005, Wang et al. 2005a, Xu et al. 2005). Matrigel is not an optimal matrix for cells because it contains many unknown components and is of non-human origin. To replace Matrigel with alternatives, several research groups have used laminin (Xu et al. 2001, Beattie et al. 2005, Genbacev et al. 2005), fibronectin (Amit et al. 2004, Noaksson et al. 2005) and human serum (Stojkovic et al. 2005a) -coated plates for hESC. Table 3 summarizes different coating materials which have been used as replacement of feeder cells in hESC cultures. A synthetic human matrix, such as recombinant human laminin (Nikolova et al. 2006), would be optimal for hESC cultures. Reproduction (2006) 132 691–698 Culture medium supplements Unlike in the mouse (Smith et al. 1988), leukemia inhibitory factor (LIF) is not able to maintain the nondifferentiated growth of hESC without feeder cells (Thomson et al. 1998) and so far no such component having an important effect on hESC as LIF has for mouse ESC has been identified. To replace feeder cells and conditioned medium in hESC cultures, identification of conditioned medium components facilitating non-differentiated growth would be crucial. Conditioned medium most likely contains variable amounts of growth factors with batch-to-batch variation. There are also differences between conditioned media from different feeder cells in supporting the growth of undifferentiated hESC (Xu et al. 2001). In our laboratory, we have not succeeded in culturing non-differentiated hESC in feeder-free cultures for longer than a few passages using conditioned medium from human foreskin fibroblasts (unpublished). Several research groups have reported successful use of supplements in hESC culture medium, such as bFGF and hyaluronic acid (Heins et al. 2004), transforming growth factor-b (TGF-b), bFGF and LIF (Amit et al. 2004), bFGF and GSK3 inhibitor (Sato et al. 2004), activin A and bFGF (Xiao et al. 2006), and spingosine-1-phosphate with platelet-derived growth factor (Pebay et al. 2005). Recently, two groups found that a combination of the BMP signaling antagonist Noggin and high concentration of bFGF was sufficient to maintain undifferentiated hESC in feeder-free culture conditions (Wang et al. 2005a, Xu et al. 2005). In our laboratory, these factors in similar high concentrations proved unsuitable to our hESC (unpublished). Beattie et al. (2005) showed that hESC cultured in the presence of activin A, nicotinamide (NIC), and keratinocyte growth factor (KGF) remained undifferentiated. They suggested that activin A may block the differentiation and NIC and KGF may have function in hESC proliferation. In addition to this, Vallier...
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This document was uploaded on 09/21/2013.

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