玻璃化冻存对胚胎神经细胞活性及脊髓移植的影响

Effect of Vitrified Cryopreservation on Embryonic Neurocyte Viability and Transplantation for Spinal Cord Injury in Rats

采用玻璃化冻存技术对大鼠胚胎神经细胞进行深低温保存，在冻存１ｄ、１０ｄ、２０ｄ和４０ｄ后，３８℃水浴快速复温，离心洗脱冷冻保护剂，检测胚胎神经细胞的活性与功能，并移植于脊髓损伤大鼠．实验结果表明：玻璃化冻存４０ｄ的胚胎神经细胞，存活率为７６．４±８．３％，膜完整性为冻存前的６９．４±７．８％．细胞活力虽有下降，但仍可维持较高水平．培养１周的部分神经元建立突触联系。

Rat embryonic cerebral neurocytes(RECN) were cryopreserved by vitrification at-196℃. The samples were separately thawed after storage for 1,10,20 and 40 days,and then cryoprotective solution was diluted and removed. RECN were assayed for their viability and function,and were transplanted for spinal cord injury in rats. The results showed that RECN cryopreserved by vitrification for 40 days retained survival rate of 76.4±8.3%. Compared with the fresh neurocytes,the cell viabiliry and membrane integrity of the frozen thawed RECN were decreased,but they still maintained at relatively high level.Synaptic connection was established in neuron culture for 1 week. Cryopreserved RECN were transplanted for spinal cord injury in rats,there was some recovery both in sensation and movement function. No obvious difference was found between vitrification and slow cooling group.