REVIEW pre Culture conditions for hESC

As a renewable source for cell transplantation for

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Unformatted text preview: renewable source for cell transplantation for severe degenerative diseases. q 2006 Society for Reproduction and Fertility ISSN 1470–1626 (paper) 1741–7899 (online) The successful differentiation of hESC to multiple cell lineages is not the topic of this review and are described elsewhere (see for example, Hoffman & Carpenter 2005). To enable the use of hESC in cell transplantation in human, it is necessary to eliminate the risk of infection transmitted by retroviruses and other animal pathogens, and immunoreactions caused by animal substances in cell cultures (Martin et al. 2005) and to establish robust and cost-effective systems for culturing hESC. This is important not only for clinical use of the cells, but also for reliable research carried out using these cells. It is difficult to evaluate the influence of various culture conditions on cells. Hence, a defined culture system with known composition of culture medium is desirable to minimize the variability that affects the reproducibility of research results. Standardized and optimized culture systems are needed, and the normality of the cells has to be followed up regularly. Derivation of human embryonic stem cell lines The first described establishment of hESC lines (Thomson et al. 1998) included the use of pronase to remove zona pellucida, and immunosurgery for isolation of the ICM DOI: 10.1530/rep.1.01079 Online version via www.reproduction-online.org 692 H Skottman and O Hovatta Table 1 Derivation methods for removal of zona pellucida and isolation of inner cell mass of blastocyst for the establishment of human embryonic stem cell line. Pronase Tyrode’s acid Immunosurgery Mechanical Reference Yes – Yes – – Yes – Yes – Yes Yes – – – Yes Yes Thomson et al. (1998); Reubinoff et al. (2000); Richards et al. (2002); Hovatta et al. (2003); Pickering et al. (2003); Mitalipova et al. (2003); Strelchenko et al. (2004); Stojkovic et al. (2004); Heins et al. (2004); Draper et al. (2004); Klimanskaya et al. (2005); Lee et al. (2005); Oh et al. (2005); Chen et al. (2005); Inzunza et al. (2005); Kim et al. (2005b); Hong-mei & Gui-an (2006); Mateizel et al. (2006); Ludwig et al. (2006) Lanzendorf et al. (2001); Cowan et al. (2004) Strelchenko et al. (2004); Kim et al. (2005b); Wang et al. (2005c) Genbacev et al. (2005) from the blastocyst. Several research groups have since published the derivation of new hESC lines using similar ICM isolation protocols (Table 1). However, immunosurgery may not be the optimal derivation method when poor quality discarded embryos with hardly visible ICM is used (Kim et al. 2005a). Immunosurgery also involves animal-derived substances, mouse antibodies, and guinea pig complement, which are not desirable thinking of cell transplantation. For standardization of derivation methods, more simplified protocols would be appreciated. Tyrode’s acid for the removal of zona pellucida and mechanical isolation of ICM is probably advantageous thinking of the...
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