摘要
在 DMEM中添加 10 %小牛血清 (NBS) ,并分别添加不同浓度的抗冻剂二甲基亚砜 (DMSO)、丙二醇 (PG)、乙二醇 (EG)及甘油 (G) ,对 7日龄小鼠生精上皮单细胞进行冷冻保存 ,复苏后台盼蓝染色测定细胞复苏率。结果 :当冻存液中 DMSO分别为 5 %、10 %、15 %、2 0 %、2 5 %时 ,细胞复苏率分别为 77.1%、88.2 %、86 .5 %、73.5 %、6 5 .5 % ,其中10 %和 15 % DMSO组细胞复苏率最高 ,与其余各组间均差异极显著 (P<0 .0 1)。当冻存液中 PG分别为 5 %、10 %、15 %、2 0 %、2 5 %时 ,细胞复苏率分别为 6 6 .2 %、84 .3%、72 .1%、6 9.9%、4 7.5 % ,其中 10 % PG组细胞复苏率最高 ,与其余各组间均差异极显著 (P<0 .0 1)。当冻存液中 EG分别为 5 %、10 %、15 %、2 0 %时 ,细胞复苏率分别为 6 4 .9%、81.6 %、6 0 .9%、4 4 .7% ,其中 10 % EG组细胞复苏率最高 ,与其余各组间均差异极显著 (P<0 .0 1)。当冻存液中 G分别为 5 %、10 %、15 %、2 0 %、2 5 %时 ,细胞腹苏率均低于 10 %。 10 % DMSO组细胞复苏率与 10 % PG组和 10 % EG组之间均差异显著 (P<0 .0 5 )或极显著 (P<0 .0 1)。结果表明 ,10 % DMSO、10 % PG及 10 % EG均适宜冷冻保存小鼠精原细胞 ,以 10 % DMSO的冻存效果最好 ,而 G则不适宜冷冻保存。在含 10 %
The single cells from 7 day old mouse seminiferous epithelia were cryopreserved,and the cell recovery rates were examined by trypan blue exclusion staining after thawing.The freezing solutions were DMEM/10% NBS supplemented with dimethyl sulphoxide(DMSO),propanediol(PG),ethylene glycol(EG) and glycerol(G) at concentrations of 5%,10%,15%,20% and 25% respectively.The cell recovery rates of DMSO at concentrations ranging from 5% to 25% were 77 1%,88 2%,86 5%,73 5% and 65 5% respectively.There were significant differences between the cell recovery rates of DMSO at 10% concentration and at other concentrations.The cell recovery rates of PG at concentrations ranging from 5% to 25% were 66 2%,84 3%,72 1%,69 9% and 47 5% respectively.There were significant differences between the cell recovery rate of PG at 10% concentration and at other concentrations.The cell recovery rates of EG at concentrations ranging from 5% to 20% were 64 9%,81 6%,60 9% and 44 7% respectively.There were significant differences between the cell recovery rate of EG at 10% concentration and at other concentrations.The cell recovery rates of G at concentrations ranging from 5% to 25% were all below 10%.The highest cell recovery rates of the four cryoprotectants showed difference or significant difference between them.The minimum cell loss of DMSO,PG,EG and G were 8 6%,12 5%,15 2% and >80% respectively.The results suggest that 10% DMSO,10% PG and 10% EG are suitable for cryopreservation of mouse spermatogonia,and 10% DMSO is the best,while G is unsuitable.The two step freezing in 10% DMSO cryopreservative solution,liquid nitrogen storing and 37℃ water bath thawing would be considered a cryopreservative protocol with high cell recovery rate for mouse sprmatogonia.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2002年第1期94-95,共2页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目 (3 9770 5 5 4)
关键词
抗冻剂
冷冻保存
生精上皮
精原细胞
小鼠
cryoprotectant
cryopreservation
seminiferous epithelium
spermatogonium
mouse
