不同冻存液对人羊膜间充质干细胞-80℃冻存与复苏效果的影响

Effects of different culture-medium on cryopreservate and resuscit of human amnion mesenchyme stem cells at-80 ℃

目的:观察不同冻存液对人羊膜间充质干细胞冻存复苏后的存活率及再培养情况的影响,并探索简单有效的冻存与复苏方法。方法:将含有血清培养基短期体外培养的人羊膜间充质干细胞分为4组,分别加入不同冻存液(体积分数):A组10%二甲基亚砜+40%胎牛血清+50%DMEM/F12,B组10%二甲基亚砜+90%胎牛血清,C组5%二甲基亚砜+4.5%羟乙基淀粉+20%胎牛血清+70.5%DMEM/F12,D组10%胎牛血清+90%DMEM/F12。以相同方法冻存并复苏细胞,比较4组细胞回收率及再培养情况。结果:A、C2组细胞复苏后回收率高于另外2组(P均<0.05),且于复苏后再培养24h内出现较多贴壁细胞。结论:以5%二甲基亚砜+4.5%羟乙基淀粉+20%胎牛血清+70.5%DMEM/F12作冻存液冻存羊膜间充质干细胞效果好。

Aim：To research the living rate of amnion mesenchyme stem cell （ amnion MSC） and the status of the cells＇ re-culture after cryopreservation and resuscitation, to probe the simple and effective way of cryopreservation and resuscitation. Methods：Amnion MSC which short-term cultured by blood serum culture-medium was mixed with four different solution for cryopreservation to be cryopreservated and resuscited in same way. The culture-medium was 10% DMSO ＋ 40% FBS＋50% DMEM/F12（group A）,10% DMSO＋90% FBS （group B）, 5% DMSO＋4.5% HES＋20% FBS＋70.5% DMEM/F12（group C）, and 10% FBS ＋ 90% DMEM/F12 （group D）. The living rate and re-culture state were compared. Results ： The living rates of group A and C were significantly higher than those of group B and D （P 〈 0.05）. And more adherence cells appeared in 24 hours after resuscitation. Conclusion： cryopreservate amnion MSCs with 5% DMSO ＋ 4.5% HES ＋20% FBS could get the best effect.