大鼠胚胎神经干细胞的冷冻复苏

目的：建立大鼠胚胎神经干细胞冷冻复苏方法，探讨冻存后神经干细胞的活力及生物学特性。方法：采用10％BSA＋7．5％DMSO作冷冻保护剂，于液氮中冻存；采用细胞培养和间接免疫荧光染色方法对冻存后细胞的活力，形态及分化能力作鉴定。结果：不同的冻存时间，细胞代数，组织来源以及神经营养因子对冻存后细胞存活率没有明显差异（P＞0．05）。冷冻保存复苏后培养的大鼠胚胎神经干细胞能够存活，能在体外多次传代，并能分化成神经元，星形胶质细胞细胞和少突胶质细胞。结论：大鼠神经干细胞的冻存复苏并不影响其原有的生物学特性包括其正常的，增殖能力和多向分化能力。

Cryopreservation of neural stem cells derived from embryonic rats

Objective:To establish a cryopreservation method for neural stem cells (NSC) obtained from embryonic rats and to study the viability and differentiation of these cells after cryopreservation. Methods:Stem cells in 10%BSA+7.5%DMSO were stored in liquid nitrogen. Cell culture and indirect immunofluorescence staining were used to identify the viability, morphology and differentiation of neural stem cells after cryopreservation. Results:Different time of cryopreservation, different passages, different sources of tissue and different trophins did not affect NSC survival ( P >0.05). After cryopreservation, embryonic neural stem cells could survive and expand in vitro for many passages, and differentiate into neurons, astrocytes and oligodendrocytes. Conclusion:Cryopreservation and resuscitation did not change biological properties of embryonic NSCs, including their morphology and capacity for expansion and differentiation. [