摘要
为了提供快速冷冻前胚胎发生部分脱水的科学依据，我们观察了家兔桑椹胚在冷冻保护液中的体积变化。当桑椹胚置于仅含３．０ｍｏｌ／Ｌ或４．０ｍｏｌ／Ｌ甘油液中，桑椹胚立即皱缩，２ｍｉｎ时体积为等渗时体积的５８．９％或５９．６％，１０ｍｉｎ时桑椹胚体积达到等渗时的９０．８％或７６．９％，将这些桑椹胚接着移入含３．０ｍｏｌ／Ｌ或４．０ｍｏｌ／Ｌ甘油加０．２５ｍｏｌ／Ｌ蔗糖保护液中，桑椹胚再次发生皱缩及膨大变化，１０ｍｉｎ后，桑椹胚体积为等渗体积的８０．８％或８１．９％。在这种脱水状态下将桑椹胚进行超快速冻存，解冻后有７３．６８％或８０．５６％在体外发育至囊胚。将经３．０ｍｏｌ／Ｌ甘油及３．０ｍｏｌ／Ｌ甘油加０．２５ｍｏｌ／Ｌ蔗糖预处理的小鼠桑椹胚超快速冻存后，有９４．１％可在体外发育。结果表明：冷冻前桑椹胚的脱水状态有助于超快速冻存的成功进行。
In order to provide mechanism of dehydration of embryos before ultrarapid freezing,we have observed the volume changes of rabbit morulae in the cryopreservation solution.When rabbit morulae were exposed to 3 0mol/L or 4 0mol/L glycerol solution at 20℃,the morulae were rapidly shrunk,the volume of the morulae was 58 9%or 59 6% of the isoosmotic volume after exposure for 2 min.At the 10th min,the volume of morulae was 90 8% or 76 9% of the isoosmotic volume.The morulae that treated by glycerol were transferred again to individual cryo preservation solutions consisting of 3 0 mol/L or 4 0mol/L glycerol plus 0 25 mol/L sucrose for 10min,they still showed firstly shrank and then swelled and reached 80 8% or 81 9% of isoosmotic volume.Under this dehydration condition,73 68% and 80 56% of rabbit morulae developed to blastocysts in vitro after ultrarapid freezing.The development rate of mouse morulae,which were pretreated with 3 0 mol/L glycerol and thereafter with 3 0mol/L glycerol plus 0 25mol/L sucrose,was 94 1% after ultrarapid freezing.These results indicated that the dehydration conditions of morulae were useful to sucessful cryopreservation by ultrarapid freezing.
出处
《解剖学报》
CAS
CSCD
北大核心
1998年第3期307-311,I018,共6页
Acta Anatomica Sinica
关键词
哺乳动物
桑椹胚
甘油蔗糖液
渗透反应
冻存
Mammalian morulae
Glycerol sucrose solution
Osmotic response
Ultrarapid freezing
