中国红豆杉悬浮培养细胞的超低温保存

Cryo-preservation of Taxus chinensis cell suspension cultures

对中国红豆杉悬浮细胞超低温保存中几个主要因素进行多方面对比研究。结果表明 ,取培养 16d的细胞进行超低温保存效果最好 ;10 %DMSO +8%葡萄糖作为冰冻保护剂对冷冻细胞起到最佳的保护效果 ;较好的降温程序是在 0℃中预处理30min后移入 - 2 0℃中停留 180min ,然后转入 - 70℃中停留 30min ,最后投入 - 196℃液氮中保存。该实验还对保存后细胞的恢复性生长进行了验证。

In this preliminary experiment some related factors for germplasm cryo-preservation of Taxus chinensis cell cultures were studied.The result showed that cell cultures should taken at 16 day after subculture for cryo-preservation.10% DMSO+8% glucose was the optimal cryoprotectants.For the appropriate freezing procedure,the cells were pre-treated at 0℃ for 30 minutes,then kept -20℃ for 180 minutes and -70℃ for 30 minutes,at last put into liquid mitrogen(-196℃) for preservation.The experiment indicated it is possible to use this freezing method for germplasm cryo-preservation of Taxus chinensis cell cultures.