摘要
目的:研究胚胎大鼠神经干细胞(NSCs)冻存复苏后的存活率及再培养生长情况,观察低温冻存对NSCs生物特性的影响。方法:含有10%BSA及7.5%DMSO的NSCs无血清培养液(不含神经生长因子)作为冻存液,将利用无血清培养液体外培养的原代、第3代、第6代胚胎大鼠NSCs进行冻存,于1周、4周、8周、12周、16周分别复苏,计数活细胞比例并进行再培养、分化鉴定。结果:不同的细胞代数,不同的冻存时间对复苏后的细胞存活率都无明显影响。复苏后NSCs的存活率均达到60%～70%,并能分化为神经元和星型胶质细胞。结论:本实验成功地对胚胎大鼠NSCs进行了冻存复苏及再培养,为择期应用NSCs耳蜗内移植治疗感音神经性聋的实验研究奠定了基础。
Objective：To explore the survival rate of neural stem cells（NSCs） from fetal rat and its biological properties after cryopreservation and thawing.Method：Different generations NSCs （the first generation,the third generation and the sixth generation）from fetal rat which cultivated with serum-free medium in vitro were cryopreserved in the cryogen, which were neurosphere culture medium with 10%BSA and 7.5%DMSO（without neural growth factor）.The cryopreserved cells were resuscitated at 1st week, 4th week, 8th week,12th week and 16th week respectively. The survival rate of cells were calculated and the cells were incubated and differentiated again.Result：Different time of cryopreservation,different generations did not affect NSCs survival（P〉0.05）after cryopreservation .The survival rate of NSCs was from 60% to 70% after resuscitated, which were differentiated into neurons and astrocytes in 10% embryonic bovine serum（without growth factor）.Conclusion：NSCs were successfully cryopreserved, resuscitated and recultured, which would create bases for the experimental study on the selected-date application of NSCs transplantation into cochlea for treating sensorineural deafness.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2010年第7期311-314,共4页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
基金
浙江省自然科学基金(No:Y2080334)
浙江省医药卫生科学研究基金计划(No:2008A056)
关键词
听觉丧失
感音神经性
神经干细胞
胚胎
冻存
复苏
hearing loss
sensorineural
neural stem cells
fetal
cryopreserve
resuscitate
