大鼠胚脑细胞玻璃化冷冻保存研究

Study on the vitrification of rat embryonic cerebral cells.

在2 0℃室温下,用玻璃化冻存液(含5 .5 m ol/ L乙二醇和1m ol/ L蔗糖)对大鼠胚脑细胞进行一步法和两步法深低温保存.在冻存1、3、7、15 d后,37℃水浴快速复温,洗脱冻存保护剂,检测细胞活性.结果表明:两步法0 .5 min平衡组,在玻璃化冻存15 d后,胚脑细胞的存活率和SDH活性分别为(78.6±3.3) %和0 .6 9±0 .0 5 ,细胞活性水平最高.此外,一步法1m in平衡组,冻存15 d后,细胞活性也较高,细胞存活率和SDH活性分别为(77.9±4 .7) %和0 .6 8±0 .0 4 .冻存后的胚脑细胞培养15 d后。

Rat embryonic cerebral cells(ECC) were vitrified with one-step and twe-step methods in solution at room temperature (20 ℃). The vitrification solution was made up of 5.5 mol/L ethylene glycol and 1 mol/L sucrose. The samples were thawed after storage for 1,3,7 and 15 days and then the cryoprotective solution was diluted and removed. ECC were assayed for their viability and function. The results showed that ECC vitrified by one-step method with 1 min equilibration and twe-step method with 0.5 min equilibration had the highest survival rate of (77.9±4.7)% and (78.6±3.3)%, and the corresponding vitality of SDH was 0.68±0.04 and 0 69±0.05, respectively. Cryopreserved ECC could develop and differentiate normally and synaptic connection was established in neuron culture for 15 days.