人胎儿肝干细胞异种移植治疗小鼠暴发性肝功能衰竭

Study on the Treatment Effects of Human Fetal Hepatic Stem Cell Heterotransplantation on Fulminant Hepatic Failure in Mice

目的:探讨人胎儿肝干细胞异种移植治疗暴发性肝功能衰竭伴严重联合免疫缺陷(SCID)小鼠的可能性与有效性。方法:改进Seglen胶原酶(Ⅳ型)原位灌注结合Percoll密度梯度离心法分离纯化中期妊娠流产的男性胎儿肝干细胞,经肝脏注射移植(106细胞)治疗D-氨基半乳糖(1600mg/kg)诱发的暴发性肝功能衰竭雌性SCID小鼠(治疗组),于移植前及移植后24,48,72h及1周取血测定血氨、谷丙转氨酶(ALT)、总胆红素(TBiL),于移植后1,2,4周取受体肝脏组织,PCR方法检测性别决定因子,并与未移植肝干细胞小鼠(对照组)比较。结果:治疗组小鼠中位生存时间较对照组延长(123.4hvs70.4h)(P<0.05);移植后治疗组血清ALT、TBiL及血氨水平降低,各时间点均低于对照组(P<0.05);治疗组肝脏病理损伤逐渐减轻,存活2周小鼠肝脏组织中性别决定因子呈阳性表达,随时间延长表达增强。结论:人胎儿肝干细胞移植能延长暴发性肝功能衰竭SCID小鼠的存活时间,改善其肝功能及肝脏病理指标。

Objective： To study the feasibility and efficiency of human fetal hepatic stem cell （HSC） heterotransplantation on fulminant hepatic failure （FHF） associated with severe combined immune deficiency （SCID） in mice. Methods： Male fetal HSC from abortion fetus in the second pregnancy trimester were isolated and purified by eollagenase （type Ⅳ） digestion method and Percoll gradient centrifugation. The purified ceils were transplanted into the livers of female SCID mice with FHF induced by D-galactosamine （1 600 mg/kg） in HSC transplantation group. The blood ammonia, ALT and TBil levels were measured pre and 24, 48, 72 h and 1 week after transplantation. Liver tissues were taken that in 1, 2 and 4 weeks after transplantation, and the sex determining region was identified by PCR technique and compared with control group. Results： The median survival time in HSC transplantation group was longer than that in control group（123.4 h VS 70.4 h, P 〈 0.05）. The serum ALT, TBiL and blood ammonia levels decreased after hepatic stem cell transplantation, and lower than those in control group at each time point （P 〈 0.05）. The pathological injury of liver improved in HSC transplantation group, and the expression of sex determining region was positive in the livers of female recipient mice 2 weeks after transplantation, and the expression increased with times. Conclusion： Stem cell transplantation can prolong the survival time of SCID mice with FHF, and improve the liver function the pathological indexes.