Derivation of embryonic stem cell line from frozen human embryos and neural differentiation.

We established a human embryonic stem cell line derived from frozen human embryos of Chinese origin. The cell line expressed the pluripotent markers SSEA-4, TRA-1-60, TRA-1-81, Oct-4, and alkaline phosphatase. The pluripotency of the cell line was also demonstrated in vivo by teratoma formation in severe combined immunodeficiency mice. The embryonic stem cells formed embryoid bodies after culturing in suspension for 7 days. The embryoid bodies were transferred to an adherent culture system in serum-free medium. The differentiating cells derived from the embryoid bodies expressed Nestin and Sox2, markers of neural progenitor cells. After the induction of cyclic AMP for 7 days, the neural progenitor cells had differentiated into neurons and glial cells.