The capacity of pluripotency and self-renewal of human embryonic stem (hES) cells may provide unlimited cell source for cell replacement therapy. In this manuscript we summarize hES differentiation protocols coculture with PA6 or MS5 stromal cells. After 7∼ 9 days of differentiation, neural rosettes were robustly appeared followed by coculture with sonic hedgehog over-expressing stromal cells efficiently generated dopaminergic neural precursor cells. Using this protocol, the majority of differentiated hES cells contained nestin positive cells (＞95%) and after final differentiation a high percentage of TuJ1 positive neurons was tyrosine hydroxylase positive (＞40%).
Keywords: Dopamine neuron; Human embryonic stem cells; Stromal cells.