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A Research for cryopreservation and transportation of embryos of small lsboratory animals.

小型实验动物胚胎冷冻保存和运输的研究。

基本信息

批准号：
02558008
负责人：
KASAI Noriyuki
金额：
$ 2.62万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

We attempted to develop two embryo transport systems using nonfreezing techniques and using a combination of vitrification techniques and a non-liquid type liquid nitrogen container. For nonfreezing technique, mouse morulae were kept in 0.5M Sucrose-PB1 solution in a thermos bottle with ice-water, followed by packaging with polystyrene foam. For vitrification technique, mouse morulae were kept in 40% ethylene glycol, 30% Ficoll and 0.5M sucrose in a 0.25ml plastic straw, followd by immediate immersing in liquid nitrogen and packaging in a non-liquid type ultralow temperature container with a special shipping enclosure. The packages were transported from Kochi to Sapporo, about 1300km by a forwarding agent. All packages arrived at the destination in 48 hours. After shipping, the survival of embryos was 46.8% for nonfreezing technique and 88.0% for vitrification techniques when evaluated by development to blastocysts in vitro. A final birthrate was 14.1% and 45.8% for nonfreezing techniques and vitrification techniques, respectively. Both systems are cheap, convenient and practical for embryo transports, and can be selected by the situation of sending and receiving facilities.We also attempted to improved the preservation technique by vitrification for mouse embryos and rat embryos. These improved preservation techniques are very simple and much helpful for prevailling embryo preservation and transport system.We also showed embryo transfer technique can clear up of infections organisms from conventional animals, indicating that we may disregard the difference of microbiogical level between the donor and recipient laboratories owing to the intrinsic aseptic condition of treated embryos.

我们试图开发两种胚胎运输系统，使用非冷冻技术和使用玻璃化技术和非液体型液氮容器的组合。对于非冷冻技术，将小鼠桑椹胚保存在装有冰水的热水瓶中的0.5M蔗糖-PB 1溶液中，然后用聚苯乙烯泡沫包装。对于玻璃化冷冻技术，将小鼠桑椹胚保存在0.25ml塑料吸管中的40%乙二醇、30%Ficoll和0.5M蔗糖中，随后立即浸入液氮中并包装在具有特殊运输外壳的非液体型超低温容器中。这些包裹是由一家货运代理从高知运到札幌的，全程约1300公里。所有包裹在48小时内到达目的地。胚胎运输后，以体外发育为囊胚评价，不冻存技术的存活率为46.8%，玻璃化冷冻技术的存活率为88.0%。最终出生率分别为14.1%和45.8%的非冷冻技术和玻璃化技术。这两种系统都是廉价、方便、实用的胚胎运输系统，可根据收发设备的情况进行选择，并对小鼠胚胎和大鼠胚胎的玻璃化冷冻保存技术进行了改进。这些改进的保存技术非常简单，对完善胚胎保存和运输系统有很大帮助，同时也表明胚胎移植技术可以清除常规动物的感染性微生物，表明由于处理胚胎的内在无菌条件，可以忽略供体和受体实验室之间的微生物水平差异。