A SIMPLE METHOD FOR MOUSE EMBRYO CRYOPRESERVATION IN A LOW TOXICITY VITRIFICATION SOLUTION, WITHOUT APPRECIABLE LOSS OF VIABILITY
A SIMPLE METHOD FOR MOUSE EMBRYO CRYOPRESERVATION IN A LOW TOXICITY VITRIFICATION SOLUTION, WITHOUT APPRECIABLE LOSS OF VIABILITY
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DOI：
10.1530/jrf.0.0890091
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发表时间：
1990-05-01
期刊：
影响因子：
--
通讯作者：
MACHIDA, T
中科院分区：
文献类型：
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作者：
KASAI, M;KOMI, JH;MACHIDA, T
将小鼠桑葚样暴露于含有30-50%的渗透剂（乙二醇、甘油、丙二醇）的溶液中，并置于改性磷酸盐缓冲盐水（PB1培养基）中，温度为20度。加热20分钟。在暴露于30%和40%的乙二醇（分别为98%和84%）或30%的甘油（88%）后，它们在培养中有很大比例发育成膨大囊胚。用PB1培养基或含30% Ficoll或30% Ficoll+0.5 m蔗糖的PB1将乙二醇和甘油稀释至30%和40%，浸泡在吸管中的液氮中，20度加热。C水。含有40%含菲柯尔的渗透性剂的溶液在冷却或加热时不结晶。将小鼠样品暴露于含30% Ficoll （EF）的PB1培养基中40%乙二醇或PB1培养基+30% Ficoll+0.5 m -蔗糖（EFS）中5-20分钟，温度为20度。C. EFS溶液在暴露5分钟内对胚胎无毒。当胚胎，在20度的EFS溶液中平衡2或5分钟。C，在-196度玻璃化。几乎所有的胚胎在培养中发育（97-98%），51%的胚胎在移植后足月发育成活的幼体。这种方法几乎不会导致胚胎活力的下降，可能是胚胎保存的实际用途。
Mouse morulae were exposed to solutions containing 30-50% of permeable agents (ethylene glycol, glycerol, propylene glycol) in modified phosphate-buffered saline (PB1 medium) at 20.degree. C for 20 min. A high percentage of them developed to expanded blastocysts in culture, after exposure to 30% and 40% ethylene glycol (98 and 84%, respectively), or 30% glycerol (88%). Ethylene glycol and glycerol were diluted to 30 and 40% with PB1 medium or with PB1 containing 30% Ficoll or 30% Ficoll+0.5 M-sucrose, immersed in liquid nitrogen in straws and warmed in 20.degree. C water. Solutions containing 40% of a permeable agent with Ficoll did not crystallize during cooling or warming. Mouse morulae were exposed to 40% ethylene glycol in PB1 medium containing 30% Ficoll (EF) or PB1 medium+30% Ficoll+0.5 M-sucrose (EFS) for 5-20 min at 20.degree. C. EFS solution was non-toxic to the embryos during 5 min of exposure. When embryos, equilibrated in EFS solution for 2 or 5 min at 20.degree. C, were vitrified at -196.degree. C and were warmed rapidly, nearly all embryos developed in culture (97-98%), and 51% developed to live young at term after transfer. This method, which results in virtually no decrease in embryonic viability, may be of practical use for embryo preservation.