Embryonic Rat Neural Stem Cells In Vitro Differentiation And Identification Of Experimental Research

Objective①To establish the methods of in vitro culture and identification of neural stem cells isolated from embryo rats.②To approach the characteristic of the proliferation and differentiation of neural stem cells cultured in vitro , as a fundamental research for neural stem cell transplantation.Methods①Neural stem cells were isolated from embryo rats, cultured and expanded by neurosphere formation using serum-free medium containing growth factors,bFGF and/or EGF,add B27.The cells were passaged continuously by disassociating mechanically in order to purify and to obtain the cells in bulk.obseve the cells morphous under inverted phase contrast microscope. Application growth curve estimate the capability of the cells.Immunocytochemistry was performed to detect the expression of Nestin,β-tubulin-Ⅲ, glial fibrillary acidic protein (GFAP).②The effects of bFGF and EGF on the proliferation of neural stem cells were examined by cell counting , alternative the optimization concentration of bFGF and EGF.③Add fetal bovine serum to serum-free medium,the cells were derivated and differentiate into neurons, astrocytes .Results①The proliferating cells isolated from neonatal rat brain grew into floating neurospheres in the serum-free medium with the presence of bFGF and/or EGF.the neurospheres could proliferate and passage...