不同直径神经干细胞球慢速低温冻存的研究

为提高神经干细胞冻存复苏后的效果及存活率,对不同尺寸与状态的神经干细胞球进行了在不同浓度DMSO(二甲基亚砜)下的慢速冻存比较.首先用CCK-8试剂盒测定神经干细胞的生长曲线,然后对处于对数生长期的3类神经干细胞(单细胞悬液,直径30～50 μm球,直径80～100 μm球)分别进行了7个不同浓度(3%,5%,7%,8%,10%,15%,20%)DMSO的冻存实验比较,并对冻后复苏的细胞进行活性检测和诱导分化.结果表明, 直径80～100 μm的神经干细胞球,DMSO浓度8%,复苏后细胞活率82.9%,且仍具有多项分化潜能.神经干细胞间的亲密联系和刻痕信号与直径尺寸共同作用,影响着神经干细胞的冻存复苏效果.

Slow Freezing Cryopreservation of Neural Stem Cell Spheres with Different Diameter

This study aims to improve the survival rate of the cryopreserved neural stem cell (NSC) spheres with different diameters by slow freezing with different DMSO concentrations.After measuring the growth curve by CCK-8 kit, three sorts of NSCs states in log phase (single cell suspension, NSC spheres with diameters of 30～50 μm and 80～100 μm) were respectively cryopreserved by seven concentrations of DMSO (3%, 5%, 7%, 8%, 10%, 15% and 20%) with the freezing rate of 1℃/min.Then the post-thaw NSCs were tested for the survival rate and the differentiation ability.The best result was that NSC sphere with diameters of 80～100 μm and DMSO concentration of 8% achieved the survival rate of 82.89% while remaining the multi-differentiation potentiality.The subtle cooperation of NSCs and sphere diameters affect the survival rate together.