山羊乳腺组织块冷冻保存试验

The research of cryopreservation goat mammary tissue

采用快速冷冻方法冻存山羊乳腺组织块,比较不同大小组织块、不同冻前预处理和每管冻存不同块数对冻存效果的影响。结果表明,冻前预处理0,6和12h,直径<0.3mm组织块的贴壁存活率分别为63.0%,89.8%和81.5%,直径为0.3～0.5mm的贴壁存活率分别为47.2%,63.0%和56.5%,直径为0.5～1.0mm的贴壁存活率分别为27.8%,35.2%和29.6%;每管冻存6块、9块和12块对冻存效果影响不显著,以6块最佳。结果显示,以添加100mL/LDMSO的D-MEM/F12为冷冻液,将直径<0.3mm的组织块冻前预处理6h,山羊乳腺组织块的保存效果较好。

<Abstrcat>Various tissue sizes,different recovered times and varied numbers of tissue fragments in each vial were analyzed using overspeed method of cryopreservation of goat mammary tissue.With the diameter of tissue fragments less than 0.3 mm,the recovered times 0,6 and 12 hours,their plating efficencies after thawing were 63.0%,89.8% and 81.5% respectively;With the diameter of tissue fragments between 0.3 mm to 0.5 mm,the recovered times 0,6 and 12 hours,their plating efficencies after thawing were 47.2%,(63.0%) and 56.5% respectively;With the diameter of tissue more than 0.5 mm,the recovered times 0,6 and 12 hours,their efficencies after thawing were 27.8%,35.2% and 29.6% respectively.However,6 masses,9 masses and 12 masses per vial respectively had no remarkable effect on the plating efficency,and 6 masses per vial was the best.The study showed,with 100 mL/L DMSO added to D-MEM/F12 as cryoprotectants,6 hours' equilibration of the diameter of tissue fragment less than 0.3 mm before freezing had better effect on the freezing of goat mammary tissue.