胚胎肝干细胞分离与移植治疗裸鼠急性肝损伤

Fetal liver stem cells for the treatment of murine liver injury

目的探讨胚胎肝脏原始／干细胞移植治疗急性肝功能损伤裸鼠的可能性与有效性。方法酶消化法分离孕13．5d的BALB／C小鼠的胚胎肝脏原始／干细胞，免疫荧光染色法及RT—PCR法鉴定AFP、Albumin、CK19、c—Met等肝系和胆系标志物，给四氯化碳（CCl4）诱发的肝毒性损伤的裸鼠尾静脉注射4×10^5个干细胞，于注射前和注射后第1、2、4、7天取裸鼠血清，测定肝功能指标，观察肝脏组织学的改善情况。结果经分离可得到具有干细胞特点的胚肝原始细胞，治疗组裸鼠在干细胞移植后肝功能指标逐渐降低，第1、2、4天均低于对照组，治疗组裸鼠肝脏组织学的病理损伤的恢复比对照组快。结论酶消化法能够分离得到较多胚胎肝脏原始／干细胞，并且具有一定的改善急性肝损伤裸鼠的肝功能及肝脏病理指标的作用。

Objective To isolate and indentify fetal hepatic progenitor/stem cells, and study the feasibility and effectiveness of their transplantation on acute liver injury in nude mice. Methods The primitive cells isolated from 13.5dpc pregnant mouse fetal liver by way of enzyme digesting were cultured in vitro and liver specific markers as AFP, CK19, Albumin, c-Met, were identified by immunofluorescence and RT-PCR on colonies. 4 × 10^5 cells were transplanted into nude mice with carbon tetrachloride induced acute liver injury. Hapatic functions were measured pre- and -post transplantation on days 1,2,4, 7. Meanwhile, hepatic pathology was studied. Results Compared to control group the hepatic functions gradually recovered in transplant group, on days 1,2,4 after fetal hepatic stem cell transplantation. The hepatic pathology significantly improved in stem cells transplantation group. Conclution Fetal hepatic progenitor/stem cell were successfully yielded by enzyme digest. Stem cells transplantation improved the hepatic function and pathology in acute hapatic injury model of nude mice.