RESUMO
In order to observe the growth, expansion and differentiation of the cultured bone marrow stromal cells (BMSC), we isolated the BMSC from adult SD rats and cultivated them with LIF and bFGF. Then, we cultured and induced the stem cells by using retinoic acid and the culture medium confected in our lab by ourselves. We found that the BMSC could expand and generate clones when they were cultured in vitro. These cells subcultured grew rapidly and differentiated into neuron-like cells and astrocyte-like cells. The results showed that BMSC have the abilities to self renew and differentiate, thus demonstrating the culture method we used is suitable for the culture of BMSC in vitro. The bone marrow stromal cell is not difficult to obtain; it is capable of expanding and differentiating in culture. If the culture condition is appropriate, it can differentiate into neuron and astrocyte. So, it is a kind of perfect seed cells.
Assuntos
Animais , Ratos , Actiemil , Farmacologia , Células da Medula Óssea , Biologia Celular , Técnicas de Cultura de Células , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Fator 2 de Crescimento de Fibroblastos , Farmacologia , Neurônios , Biologia Celular , Projetos Piloto , Ratos Sprague-Dawley , Células-Tronco , Biologia Celular , Células Estromais , Biologia Celular , Tretinoína , FarmacologiaRESUMO
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.