体外培养及冻存对大鼠胚胎神经干细胞生物学特性的影响

Effect of culture in vitro and cryopreservation on biological features of embryonic rat neural stem cells

目的体外培养、冻存、移植大鼠胚胎神经干细胞（NSCs），观察细胞生物学特性有无改变。方法取孕15d的大鼠海马区细胞，体外培养；以20％牛血清白蛋白（BSA）加10％二甲基亚砜（DMSO）作冷冻保护剂，于液氮中冻存；复苏后计数活细胞数，免疫细胞化学鉴定其分化状态；移植入C6大鼠胶质瘤模型观察其存活情况。结果第1-4代NSCs复苏后神经干细胞存活率在40％-63％之间，差异无显著性意义。冻存复苏后的NSCs能够增殖、传代，移植入胶质瘤模型体内能够大量存活。结论大鼠胚胎NSCs能够在体外增殖、分化。并且冻存复苏不影响其生物学特点和细胞活性。

Objective To observe the changes of biological features of embryonic rat neural stem cells （NSCs） after cultured in vitro, cryopreserved and then transferred into rat glioma. Methods The NSCs were isolated from the hippocampus of the rat embryos （fetal age of 15 d） and cultured in vitro. The NSCs at P 1-P4 were stored in the liquid nitrogen with 20% bovine serum albumin and 10% dimethyl sulphoxide. The livability and differentiation of the NSCs after resuscitated were detected by immunocytochemical staining, and the livability of NSCs transplanted into the glioma models was also observed. Results The livability was not affected by cryopreservation and the livability of the NSCs at P1-P4 was between 40% and 63%. After cryopreservation, NSCs could still survive and proliferate in vitro through several approaches, and they could also survive in the glioma models. Conclusion The biological characteristics of embryonic rat NSCs would not be affected by cryopreservation and resuscitation.