CAJ | 학술논문

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.
시험탐색료일반실험실조건하의남개현부세포초저온보존구유교고TTC검측활성적간편방법.결과표명:선택계대후2 d적의남개현부세포,세조1차,세포침정중현부우6%자당배양기,우4℃저온하단련6 h;농축세포사지유교고적세포밀도(0.25～0.30 g/mL),가입160 g/L감유작위빙동보호제,직접치우-80℃심랭빙상보존;사용시이37℃쾌속해동,리심후제거보호제,연후용거리자수세조후진행TTC활성검측,보존30 d후적세포상대활성체0.8좌우,가이만족실험실일반연구용.
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.