拟胚体对成体细胞参与胚胎嵌合的影响

The Effect on Chimeric Embryo Construction from Fibroblasts and Early Embryos about Mouse Embryoid Body

试验尝试构建成体细胞同小鼠早期胚胎的嵌合共生胚胎。取成年人皮肤组织的成纤维细胞,慢病毒转染皮肤成纤维细胞标记EGFP荧光蛋白,同小鼠早期8-细胞胚胎进行嵌合。结果显示慢病毒高效标记人皮肤成纤维细胞表达EGFP荧光蛋白,通过皮肤成纤维细胞与小鼠胚胎干细胞形成的拟胚体环境作用后,皮肤成纤维细胞成功与小鼠胚胎形成嵌合胚,嵌合囊胚形成率为38.08%,表达EGFP荧光蛋白的皮肤成纤维细胞能够嵌合到小鼠胚胎的不同部位。嵌合胚在胚胎干细胞分离培养环境下进行培养,嵌合到小鼠内细胞团的皮肤成纤维细胞参与小鼠内细胞团的组成,参与小鼠内细胞团组成的胚胎占嵌合胚比率为1.74%。小鼠胚胎干细胞拟胚体环境作用可以成功介导人皮肤成纤维细胞同小鼠早期胚胎形成嵌合共生体系。

Chimeric embryos from mouse embryos and human fibroblasts were constructed. The fibroblasts from adult foreskin were isolated, cultured and transfected EGFP with lentivirus vectors, and then chimeric embryos were constructed with mouse embryos and fibroblasts. The results showed that human fibroblasts efficiently expressed EGFP after transfected with lentivirus vectors. 38.08 % chimeric embryos were constructed successfully with mouse embryos and EGFP fibroblasts which were from embryoid bodies of mouse embryonic stem ceils and EGFP fibroblasts. EGFP fibroblasts were taken part in different ingredient of chimeric embryos. Under the circumstance of embryonic stem cell isolation and culture, EGFP fibroblasts in 1.74 % chimeric embryos became themember of inner cell masses on feeder cells. Human fibroblasts can become chimeric embryos with mouse embryos by the microenvironment of mouse embryoid bodies.