Neural stem cells transplantation is the research hotspot for the treatment of central nervous system damage and refractory disease. How to improve the survival rate and the cloning formation rate of neural stem cells will be an important foundation and prerequisite for its application.OBJECTIVE:
To study the influencing factors on the culture of neural stem cells in vitro in order to provide reference for the basic and clinical application of neural stem cells.METHODS:
Neural stem cells were isolated from the brain of Sprague-Dawley rats within 24 hours after birth, centrifuged, and then inoculated at the concentration of 1×108/L. Then the cells were pre-cultured with dulbecco's modified eagle's medium/F12 medium containing 10% fetal bovine serum for 4 hours and then serum-free conditioned medium (dulbecco's modified eagle's medium /F12, basic fibroblast growth factor, epidermal growth factor and B27) was used. The medium was changed every 2 or 3 days, and cells were passaged at 6 days. Morphological and biological characteristics of the cells were observed under inverted microscope to indentify neural stem cells. Immunocytochemistry was used to examine nestin expression on passage 3 neural stem cells and the expression of neuron-specific enolase and glial fibrillary acidic protein in differentiated cells.RESULTS AND CONCLUSION:
Neural stem cells could be obtained from newborn rats 24 hours after birth. Pre-culture with serum could increase the proliferation rate and survival rate. Immunohistochemistry combined with serum-induced differentiation could be used to identify neural stem cells. Sampling time, cell density, passage time, cytokines and serum all can influence the proliferation and differentiation of neural stem cells.RESULTS AND CONCLUSION:
Xu FC, Zou LL, Mei XM, Guo Y. Culture of neural stem cells and the influencing factors. Zhongguo Zuzhi Gongcheng Yanjiu. 2013;17(10):1835-1840.