Development of simple storage methods for donor cells such as freeze-drying for bovine somatic cell nuclear tramsfer

研究了用于体细胞核移植的供细胞的简单储存方法。首先，我们研究了是否可以从简单的冷冻组织中提取活细胞。牛的几种组织分别在-20℃、-80℃和-196℃冷冻。组织解冻后，即使在-80℃冷冻6个月后也能从耳组织中获得活细胞。然后，观察冷冻干燥后体细胞的核状态。将海藻糖引入洋地黄苷提取的耳组织细胞中，然后冷冻干燥细胞。核DNA的断裂用彗星测定法检测。0或20&#61549；g/ml洋地黄苷和0.4M海藻糖处理后的冻干细胞DNA断裂与未处理的活细胞相似。

Simple storage methods for donor cells used for somatic cell nuclear transfer were examined. First, we examined whether live cells can be retrieved from simple frozen tissues. Bovine several tissues were frozen at -20 oC, -80 oC and -196 oC. After thawing tissues, live cells were obtained from ear tissues frozen at -80 oC even after 6 months. Next, nuclear status of somatic cells after freeze-drying was examined. Trehalose was introduced into cells taken from ear tissues by digitonin, then the cells were freeze-dried. Fragmentation of nuclear DNA was examined by a Comet assay method. DNA fragmentation of freeze-dried cells treated with 0 or 20g/ml digitonin and 0.4M trehalose was similar to that of untreated live cells.