体外培养枫径猪胚胎成纤维细胞系方法的建立

Establishment of A Method for Fengjing Porcine Fetal Fibroblast Cell Line in Vitro

选取枫径猪胚胎,采用胶原酶消化培养法制备猪胚胎成纤维细胞,经数次传代培养及冷冻复苏试验表明胚胎成纤维细胞仍具有正常的传代能力。在体外培养至15代后成纤维细胞核型正常,符合体细胞克隆转基因的要求。用PCR方法建立了枫径猪胚胎成纤维细胞性别鉴定的反应体系,对其中6株细胞进行了性别鉴定。

Fengjing porcine fetal fibroblasts were cultured by fetal tissue pieces which were digested with collagenase, and an ideal cellular culture method was established. After several pasages of culture and freeze revival, the fetal fibroblast cells still had normal capacity of passage. Karyotype analysis indicated that after 15 passages of in vitro culture they showed no abnormal appearance. It suits for the need of cloning and transgenic construction. After establishing the technique of sex determination of fetal fibroblast of porcine, we identified the sex of 6 cell lines.