摘要
通过实验确定分离小鼠胚胎成纤维细胞的最适胚龄，探讨小鼠胚胎成纤维细胞饲养层的效果及其寿命。结果表明，从１０ 、１１ ｄ 小鼠胚胎所获的胚胎成纤维细胞中混有大量神经母细胞和红细胞；用１４ 、１５ ｄ 的胚胎则基本不能获得胚胎成纤维细胞；消化３ ～４ 个１２ ～１３ ｄ 胚胎所获细胞悬液可接种８～９ 个２５ ｃｍ２ 的培养瓶。从一只孕鼠的所有胚胎所获的成纤维细胞可接种约２０ 个２５ ｃｍ２ 的培养瓶。所获细胞悬液的活细胞百分率可达９０％ ，接种成活率达８５ ％ 。胚胎成纤维细胞贴壁时间为５～６ ｈ ，１２ ｈ 后进入增殖期，４８ ｈ 形成细胞单层。ＥＳ细胞克隆在小鼠胚胎成纤维细胞饲养层上形态典型、增殖迅速、不分化。饲养层寿命可维持２ ～３ 周。
The present experiment was carried out to investigate properage of the mouse embryos fitted to isolate embryonic fibroblasts andeffectiveness of embryonic fibroblast feeder layer and their life span.The results indicated:Embryonic fibrobalsts isolated from 11,12 dmouse embryosblnded with a large number of neuroblasts and erythrocytes.Embryonic fibrobalsts couldnt be isolated from 1,15 d mouse embryos.Thcell suspension digested from 3～4 embryos of 12～13 d old was enoughplanting in 8～9 25 cm 2 flasks.The suspension digested from all embryosof one pregnant mouse was enough planting in about 20 flasks of thesame size.After planting,the rate of living cells reached at 90% and the surviving rate was 85%.he time for cell attachmnt taken place was 5～6 h.After 12 h,fibroblasts began to divide and a monolayers wasformed by 48 h.Cultured on the surface of mouse embryonic fibroblastfeeder layer,ES cells clonies grew typically and fast withoutdifferentiation.The life spn of feeder layer was about 2～3 weeks.
出处
《黑龙江畜牧兽医》
CAS
北大核心
1999年第9期5-6,共2页
Heilongjiang Animal Science And veterinary Medicine
基金
国家自然科学基金
