| 摘要: |
| 采用程序化冷冻方法，对不同植冰温度下几种冷冻液的结晶变化进行了分析，并比较了其对小鼠胚胎的冷冻效果。结果表明，－5.0 ℃植冰时，3种冷冻液PEF、PGF和PEGF只有部分结晶；当植冰温度为－5.5 ℃时，PEF和PEGF完全结晶时间为10 min，PGF在15 min尚未完全结晶；－6.0 ℃植冰时，3种冷冻液完全结晶时间分别为8，10和10 min；－6.5 ℃植冰后完全结晶时间分别为5，8 和5 min。将小鼠胚胎载入上述3种冷冻液，－5.5 ℃植冰冷冻解冻后,囊胚孵化发育率分别为81.7％，68.5％和87.5％，－6.0 ℃植冰时发育率分别为80％，76.5％和90.9％，－6.5 ℃植冰后的发育率分别为10％，13.3％和38.4％。据此得出，在－5.5或－6.0 ℃时植冰,以PEGF作为冷冻保护液冷冻保存小鼠胚胎效果较好。 |
| 关键词: 植冰温度 结晶 胚胎冷冻 小鼠 |
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| 基金项目:国家“863”高技术项目(2001AA213081) |
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| Effects of crystallization of cryoprotectants at different seeding temperature on the developmental ability of mouse embryos |
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| Abstract: |
| Crystallization changes of various cryoprotectants sections were analysized at different seeding temperature using programmed freezing procedure.When seeded at -5.0 ℃，PBS was freezed completely within 2 min,PBS+ethylene glycol (PEF),PBS+glycerol (PGF) and PBS+ethylene glycol+glycerol (PEGF) were not crystallized entirely although the process lasted for 15 min;When seeding temperature was -5.5 ℃，PEF and PEGF were crystallized entirely in 10 min,but PGF was not in 15 min;While seeding temperature reduced at －6.0 ℃ and －6.5 ℃，the three kinds of cryoprotectants were solidified in 8 min,10 min and 10 min vs 5 min,8 min and 5 min respectively.Mouse embryos were exposed to the three cryopreservative compounds and seeded at -5.5 ℃ and －6.0 ℃ followed lauching into liquid nitrogen at －35 ℃,after thawing and in vitro culture,percentage of hatching embryos were 81.7%,68.5% and 87.5% vs 80%,76.5% and 90.9% ,they both had higher developmental abilities than that seeded at －6.5℃ (10%,13.3% and 38.4%).Compared with PEF and PGF,PEGF had better effect on the freezing of mice embryos. |
| Key words: seeding temperature crystallization embryo cryopreservation mouse |
