Experimental study on in vitro inducement differentiation of neural stem cells from fetal rat ependyma

Objective To explore the feasibility and regularity of neural stem cells (NSCs) culture and inducement differentiation from fetal rat ependyma, and offer the positive control for searching the NSCs seed cells outside the nerve tissue. Methods The cells of fetal rat ependyma were cultured in NSCs medium in order to ascertain the optimal survival conditions of NSCs in vitro. The involved cytokines mainly included EGF, bFGF and LIF (20 ng/ml respectively). The proliferation of NSCs was confirmed by formation of cell clones. The differentiation of NSCs was primarily estimated by observing the cellsmorphology under a microscope. Results The NSCs derived from the fetal rat ependyma were observed to rapidly proliferate to form some cellular spheres or neural spheres consisting of NSCs. Having been separated into single cells and then plated, the islets-shaped cellular spheres were found to appear again. Continuous incubation of the NSCs in NSCs-medium could lead to their further proliferation. Some of them became large enough to grow into cell-body, and some small buds were formed from several NSCs bodies and then developed into the long projects that might be connected with each other. Conclusion It is feasible for the fetal rat ependyma to be induced into NSCs. The gradual developmental process of NSCs follows the rule of growing from single NSCs, to neural sphere and at last to the differentiated neurons, through which the NSCs manifest themselves with characteristics of proliferation and differentiation different from the general neurons.