Isolation and differentiation of neural stem cells from neonatal rats in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND: Neural stem cells are always derived from fetal rats and adult rats, and it is complex to isolate the cells by cell culture.OBJECTIVE: To study a convenient and effective method for the isolation and the culture of neural stem cells in neonatal rats.DESIGN, TIME AND SETTING: An observation study based on cells was carried out in the Chongqing Medical University (Chongqing, China) from October 2006 to March 2007.MATERIALS Wistar neonatal rats of 1-3 days old.METHODS: Subsequent to trypsin digestion, primary culture of the cells was performed in serum-free suspension culture medium. Then the cells were induced to incubate in DMEM/F12 containing 0.10 volume fraction of fetal bovine serum. MAIN OUTCOME MEASURES: Phase contrast microscopy was employed to observe the growth of neural stem cells and the morphology of the differentiated cells. Neural stem cells and the differentiated neurons were identified using indirect immunofluorescence cytochemistry, as well as expression of gilal fibrillary acidic protein. Moreover, the proliferation of the BrdU-labeled neural stem cells was also investigated.RESULTS: The neural stem cells isolated from neonatal rat brains had the potential of serial passage and proliferation, besides, they express neuroepithelial stem cell protein (nestin) and differentiate into neurons, astrocytes and oligodendrocytes.CONCLUSION: Neural stem cells can be harvested from neonatal rat brains at a large scale, and they maintain their undifferentiated features and have the capacity of self-renewal and pluripotentiality.