甘草悬浮细胞的玻璃化法超低温保存研究

Studies on the Cryopreservation of Suspension Culture Cell in Glycyrrhiza in flata Batal by Vitrification Approach

以甘草悬浮细胞为材料,进行了超低温保存技术的研究。高产黄酮的甘草悬浮细胞是一种极具价值但又难于保存的材料,为解决甘草细胞超低温保存中存在的关键问题,即如何减少渗透压的突然改变对细胞造成损伤,通过优化预培养时间、细胞年龄、预处理时间、脱水时间、洗涤液蔗糖浓度等影响因素,建立了一套适合于甘草悬浮细胞的保存方法。采用改良的2,3,5-氯化三苯基四氮唑(TTC)法检测,此法存活率可达82.9%。将保存后的细胞进行恢复培养,其恢复生长率可达80.4%。

A procedure had been developed for cryopreservation of Glycyrrhiza inflata Batal suspension cultures by vitrification.High flavonoid-yield G.inflata Batal suspension cultures are valuable materials but hard to be preserved.The key point of this research is how to reduce the damage of cell caused by the sharp change of pressure.So setting up a suitable method must depend on various important factors such as the duration of regrowth,the harvest time,the duration of dehydration,the sucrose concentration of washing solution,etc.Survival rate of G.inflata Batal suspension culture cell was 82.9% by TTC examination,and regeneration rate reached to 80.4%.