Human Embryonic Stem Cells And Rabbit Embryonic Stem Cell Line Establishment And Identification

In this study, we reported seven hES cell lines (SH4, SH28, SH35, SH38, SH39, SH42 and ICSI-ES) being established from the Chinese population and characterized in a comparative way. The blastocyst used for embryonic stem cell derivation are clinically IVF surplus and ICSI-produced blastocysts. All seven cell lines had morphological characteristics typical of hES cells. The cell lines expressed alkaline phosphatase activity and molecules typical of primate embryonic stem cells, including Oct-4, Nanog, TDGF1, Sox2, EBAF, Thy-1, FGF4, Rex-1, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81. Five of the lines (SH4, SH35, SH38, SH39, and SH42) formed embryoid bodies that expressed markers of a variety of cell types (ICSI-ES in vitro differentiation has not been done); four of them (SH35, SH38, SH39, SH42) formed teratomas with tissue types representative of all three embryonic germ layers (ICSI-ES in vivo differentiation to be done). These human embryonic stem cells are capable of producing clones of undifferentiated morphology, and one of them (SH35) was propagated to become a subline (SH35a) which still retained hES cells characteristics and pluripotential differentiation capacity in vitro and in vivo. The newly established cell lines showed PD values that ranged from 30 to 39 h. At present, SH4, SH28, SH35, SH38, SH39, SH42, ICSI-ES have reached approximately 315, 310, 230, 250, 206, 230, 120 PD, respectively. After long term culture in vitro, the lines remained undifferentiated, and retained normal karyotypes of 46(with SH4, SH28, SH35, SH38 and ICSI-ES as XX, and SH39 and SH42 as XY). These results demonstrate that the cell lines can be cultured for prolonged periods without compromising their chromosomal stability.