从早期胚胎分离和培养兔胚胎干细胞

Isolation and culture of ES-like cell from rabbit preimplanted embryos

采用昆白系小鼠成纤维细胞(MEF)作为饲养层,低糖DMEM+10%NBS+10%FCS+10 ng/mL LIF+10 ng/mL SCF+0.1 mmol/L β-巯基乙醇+100 IU/mL青霉青+80 IU/mL链霉素作培养基,从培养96 h的胚胎中分离出1个兔类ES细胞集落,克隆传至4代,悬浮培养的兔类ES细胞团,可出现囊状胚。兔类ES细胞接种在衰老的饲养层上,出现滋养层细胞和上皮样细胞。

One rabbit ES-like cell line was isolated from 96 h blastocysts by culture system composed of embryonic fibroblast feeder of Kun Ming strain white mouse (MEF),medium (low glucose DMEM, 10%NBS,10%FCS,0.1 mmol/L β-mercaptoethunol,100 IU/mLstreptomycin sulfate) supplied with 10 ng/mL SCF and 10 ng/mL LIF.One rabbit ES-like cell clones was passaged to 4 times.When ES-like cells were cultured on degenerated MEF,cells formed embryonic-like body.And also they differentiated epithelial and trophoblastic cells.