机械吹打法、胰酶消化法分离新生大鼠脑组织神经干细胞效果观察.

Objective To compare the effects of mechanical isolation and pancreatin digestant isolation in separating neural stem cells (NSCs) from brain tissues of neonatal rats. Methods The cerebral cortex of newborn rals with in 3 days were isolated and cultured by mechanical and panerealin digestion, respectively. The NSCs were identified and differentiated by immunofluorescence assay. Diameter and number of cell spheres were detected on day 2, 5 and 7 of culture. The proliferation ability of the primary passage of NSCs and ihe third-passage NSCs in the two groups were compared by MTT. The third-passage NSCs were frozen and recovered and the survival rate of the cells was calculated. Results Both of these two methods could isolate NSCs from the cerebral cortex of neonatal rats, and those NSCs could be induced to differentiate into neurons and glial cells. On day 5 and 7, the NSCs' ball number and diameter which came from the mechanical isolation group were higher than those of the pancreatin digestant group in the same culture time (all P < 0.05). On day 2, 5 and 7, the prime NSCs which came from ihe mechanical isolation group had better clonal proliferation ability than those of the pancreatin digestant group in the same culture time (all P < 0.05). On day 2 and 5, ihe ihird-passage NSCs of the mechanical isolation group had better clonal proliferation ability than those of the pancreatin digestant group in the same culture time (all P <0.05). The survival rates of the third-passage NSCs of the mechanical isolation group and pancreatin digestanl group were respectively 85% and 89%, respectively ; and there was no significant difference between these two groups (P>0. 05). Conclusion Both mechanical isolation and pancreatin digestant isolation can successfully isolate NSCs. NSCs which come from the mechanical isolation group have better clonal proliferation ability than those of the pancreatin digestant group. [ABSTRACT FROM AUTHOR]