Study on Isolation and Differentiation Neural Stem Cells from Hippocampus in Neonatal Rats

MIAO Zongning; WU Weijiang; QIAN Hanguang; ZHAO Jidong

doi:10.19529/j.cnki.1672-6278.2016.04.18

Abstract：To establish the method of culture neural stem cells in the hippocampus of neonatal rats in vitro in order to find a suit-able cell source for the treatment of nervous system diseases.NSCs were isolated use accutase and mechanical separation method from hippocampus area of neonatal SD rat.The cells were cultured in completely medium and medium components include DMEM/F12 ser-um-free medium,B27,basic fibroblast growth factor and epidermal growth factor.Anti nestin immunofluorescence staining were used to identify the third generation cell after accutase enzyme digestion and subculture.Culture medium containing 10%fetal bovine serum were used to induce NSCs,NF-200 and GFAP immunofluorescence staining were used to identify the differentiation of NSCs to neu-rons and glial cell.In the hippocampus of neonatal rats,a large number of nerve cells were formed in the serum free medium,and some of the neural spheres appeared fused and adherent differentiation.The cells showed typical NSCs morphology.The expressions of nestin staining showed that most cells were positive.Nerve cell spheres could differentiate into neurons and glial fibers acidic protein expression positive cells after cultured with medium containing fetal bovine serum.The NSCs isolated from the hippocampus of neonatal rats has the ability of self-renewal and proliferation,which has the potential to differentiate into neurons and glial cells in the serum containing fetal bovine serum.