EXPERIMENT ON CULTURE, IDENTIFICATION OF NEURAL STEM CELL FROM SD RATS / 解放军医学杂志

ABSTRACT

To explore the feasibility of culture and identification of neural stem cells (NSCs) from ependyma, subventricular zone and cotex of fetal and adult rat respectively, and to make the basis for treatment of degenerative diseases with NSCs transplantation, all the seed cells derived from ependyma, subventricular zone and cotex were isolated from both fetal and adult SD rat, respectively. They were then cultured, induced to differentiation and subcultured continuously in a "CYTOKINE NSCs culture medium". Identification was carried out using Nestin, NSE and GFAP antibodies for differentiated NSCs, neuron and neuroglials, respectively. The seed cells from these four locations proliferated rapidly under some corresponding conditions, and formed "neurologic spheres", which consisted of many cells and expressed Nestin antigen. After continuous culture and subculture, NSCs might divide and proliferate further. Some NSCs buds developed processes and formed nerve fibers further, while the soma enlarged into the cells with "long processes", which connected or crisscrossed with each other, and were confirmed as neurons and neuroglias by immunocytochemistry. Seed cells from fetal rats might generate more NSCs than those from adult rats, and those from ependyma and subventricular zone produced more NSCs than those from cortex. There was no special morphological difference between ependyma NSCs and cortex NSCs. It is suggested that NSCs existed not only in ependyma and cotex of fetal SD rat, but also in the subventricular zone and cotex of adult SD rat. Fetal rat nerve tissue possesses much more NSCs than adult one.