The study of in vitro culture and differentiation of the neural stem cells from human embryonic hippocampus

Objective To study the conditions of in vitro culture and the features of differentiation of neural stem cells from human embryonic hippocampus. Methods The neural stem cells were isolated from human embryonic hippocampus. The cells were cultured in serum-free medium. Then the neurospheres formed after proliferation. Differentiation happened while the neurospheres adhered to the dish wall. The cells were labeled with BrdU for observation of cell mitosis and proliferation. The neural stem cells and the default differentiated cells were distinguished via immunohistochemisty. Results It was found that the neural stem cells from human embryonic hippocampi have the ability of proliferation. The doubling time is 3.2 days. They can be cultured for passage and differentiate toward multiple directions. BrdU labeled mitotic cells and cells in proliferation were observed. Nestin, GFAP, and Tuj-1 expressing cells were identified after adhesion and differentiation. Conclusions The neural stem cells isolated and cultured from human embryonic hippocampus have the ability of self-renewal and proliferation. They are able to differentiate into neurons and astrocytes, resembling the features of neural stem cells. This cell line will be very useful for basic and clinic research.