鸡小肠上皮干细胞的分离培养

Study on Isolation and Culture of Intestinal Epithelial Cell in Chicken Embryo

为建立鸡小肠上皮干细胞(Intestinal epithelial stem cells,IECs)体外培养体系并对其进行生物学特性研究,取18日龄鸡胚,分离小肠获得IECs,在DMEM/F12培养基中培养并观察其形态。通过传代及生长曲线的绘制分析其增殖情况,并经形态学观察及细胞免疫组织化学、RT-PCR等方法对所得到的纯化后的鸡小肠上皮干细胞进行鉴定。结果表明,鸡IECs可在体外扩增培养,目前传代至P11,纯化所得的鸡小肠上皮细胞经形态学观察及免疫荧光、RTPCR法鉴定为阳性,由此可得出结论:鸡小肠组织中可以分离出IECs,且能够在体外增殖培养。

This current study was conducted to investigate the biological characteristics of chicken IECs（ Intestinal epithelial stem cells） by isolating and culturing in vitro. IECs were isolated from small intestine of 18 day-old chicken embryos. The suitable culture medium for IECs was DMEM /F12. Then its proliferation was analyzed by subculture and the growth curve. The purified chicken intestinal epithelial stem cells were identified by morphology,immunohistochemical methods and RT-PCR. The results showed that chicken IECs can be cultured in vitro and have been passed to 11 generation. The purified IECs were positive by morphology,immunohistochemical methods and RT-PCR. To sum up,IECs can be isolated from small intestine,which can proliferate rapidly in vitro.