| 摘要: |
| 对半胚冷冻的方法和条件进行了系统研究,证明采用下述程序冷冻小鼠半胚可显著提高解冻半胚的成活率:①在含10％甘油的磷酸缓冲液中平衡35～40min;②在-6.5℃或-7℃诱发结晶,③以0.05℃/min的速度缓慢降温15min;④以0.3℃/min的速度降温至-35℃投入液氮;⑤在30～35℃的水浴中解冻;⑥在含不同浓度(6.6％,3.3％,0％)甘油和0.3mol/L蔗糖的磷酸缓冲液中平衡35～40min,去除甘油。将按上述程序冷冻—解冻的16对半胚移植后,获半胚鼠12只,其中5对为同卵双生。 |
| 关键词: 半胚冷冻,小鼠 |
| DOI： |
| 分类号: |
| 基金项目:国家教委“资助优秀年轻教师基金”项目 |
|
| Freezing of Mouse Demi-emdryos |
|
Zhang Yong Wang Jianchen
|
| Abstract: |
| The methods and conditions suitable to freezing of demi-embryos
were systematically studied。The results showed that freezing of demi-embryos
embedded with agar can improve the survival rate of frozen-thawed mouse
demi-embryos using the following procedure:(l)equilibration in PBS with 10%
glycerol for 35~40 min,(2)seeding at -6.5℃or-7℃;(3)slow cooling at
about 0.05℃/min for 15 min,(4)continuous cooling at 0.3℃/min to -35℃
at which demi-embryos with agar were plunged into liquid nitrogen;(5)thaw-
ing in water at 30~35℃;(6)remova1 of g1ycerol by equilibration in PBS
with 6.6%,3.3%,0% glycerol and o.3mol/L sucrose for 40(10,15,15)min.
Sixteen pairs of demi-embryos frozen-thawed by the above procedure were
transferred.of 8 recipients,5 became pregnant (62.5%) and produecd 12 young
(37.5%) at delivery,including 5 pairs of monozygotic twins(31.3%). |
| Key words: freezing of demi-cmbryo, mouse |
