Kiselev Sergey L.
(Vavilov Institute of General Genetics RAS, Moscow, Sl_kiselev@yahoo.com)
Recently we have established four new hESC lines from surplus blastocysts donated by couples undergoing IVF procedure. All 4 cell lines were propagated in vitro for more than 40 passages, maintained a normal karyotype and were viable after freezing and thawing. However, instability of karyotype was detected in 2 cell lines on early passages. Teratoma analysis of one cell line with abnormal karyotype resulted in appearance of invasive carcinoma-like cells on histological sections. Cell lines with abnormal karyotype were less potent even to form embryoid bodies. At the same time we were able to differentiate hESC lines with normal karyotype to different lineages. We have developed an efficient protocol of direct differentiation of hESCs to endothelial cells purified by magnetic sorting procedure. We have also studied differences in epigenetic status of the cell lines during their differentiation. Our data indicate on a diverse epigenetic profile for ESC lines and provide the evidence that in the course of in vitro differentiation ESCs undergo the same epigenetic program as differentiating cells of embryo in vivo.
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