拟南芥悬浮细胞超低温保存试验

Cryoperservation Test of Arabidopsis Suspension Cells

试验探索了一般实验室条件下拟南芥悬浮细胞超低温保存具有较高TTC检测活性的简便方法.结果表明：选择继代后2 d的拟南芥悬浮细胞,洗涤1次,细胞沉淀重悬浮于6%蔗糖培养基,于4℃低温下锻炼6 h;浓缩细胞使之有较高的细胞密度（0.25－0.30 g/mL）,加入160 g/L甘油作为冰冻保护剂,直接置于-80℃深冷冰箱保存;使用时以37℃快速解冻,离心后除去保护剂,然后用去离子水洗涤后进行TTC活性检测,保存30 d后的细胞相对活性达0.8左右,可以满足实验室一般研究用.

A series of experiments on cryoperservation of Arabidopsis suspension cells were performed,and a viable sample method was obtained.The optimized scheme is shown as follows： taking Arabidopsis suspension cells in 2 days after subculture,washing them for one time,resuspending the cell pellet in basic medium with 6% of sucrose,pretreating the suspension for 6 hours with 4℃ degree temperature,concentrating the suspension,adding 160 g·L-1 of glycerol as cryopretective agent,placing and keeping it in cryogenic refrigerator（-80 ℃）,thawing it with 37℃ degree temperature when using,then checking activity of cryopreserved suspension cells by TTC after washing or resuspending,the relative activity of cryopreserved suspension cells could reach 0.8 after 30 days.