脐血干细胞向肝细胞分化的初步研究  

Experimental Study on Differentiation of Human Cord Blood Stem Cells into Hepatocytes

作　　者：黄韬[1] 李波[1] 林浩铭[1] 覃扬[2] 

机构地区：[1]四川大学华西医院普外科,成都610041 [2]四川大学华西基础与法医学院分子生物学研究室,成都610041

出　　处：《中国普外基础与临床杂志》2007年第4期423-427,共5页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的探讨脐血干细胞体内定向诱导分化为肝细胞的可行性。方法将BALB/c裸鼠36只随机分成实验组和对照组,各18只;实验组再随机分成A、B、C3组,每组6只。采用^(60)Co治疗仪γ射线对实验组和对照组行350cGy的亚致死剂量照射,实验组A、B、C3组照射后3h内经尾静脉分别输入1.0×107个/只、2.0×107个/只和3.0×107个/只人新鲜脐血有核细胞,对照组经尾静脉注入等体积无菌生理盐水。1个月后对实验A、B组和对照组裸鼠经腹膜腔注射四氯化碳(CCl4),实验C组注射等体积生理盐水。2个月后采用免疫组化和RT-PCR方法检测裸鼠肝组织人源性CK18、CK19和人源性白蛋白(ALB)的表达。结果实验A组和实验B组裸鼠的肝组织人源性CK18、CK19和ALB表达阳性,实验B组的表达强度高于实验A组(P<0.05);而对照组和实验C组裸鼠肝组织CK18、CK19和ALB表达阴性。结论脐血在CCl4导致的肝脏损伤的特殊微环境内能分化为肝细胞,并表达白蛋白,其表达强度与输入的干细胞数量有关。Objective To investigate the feasibility of differentiating human umbilical cord blood stem cells into hepatocytes. Methods Thirty-six BALB/c nude mice were randomly divided into experimental group and control group（18 in each of the group）, and experimental group was again randomly divided into group A, B and C （six in each of the group）. The mice in experimental group and control group were exposed to 350 cGy radiation produced by ^60Co. After 3 h, karyocytes at different concentrations in the fresh human umbilical cord blood were injected into the mice in experimental group A, B, C via their tail veins, and the equal volume of normal sodium （NS） was also injected into control group via tail veins. After one month, carbon tetrachloride （CCl4） was injected into experimental group A, B and control group via abdominal cavity, and the equal volume of normal sodium was injected into experimental group C. After two months, immunohistochemistry and reverse transcriptase polymerase chain reaction （RT-PCR） were used to detect the expressions of human cytokeratin-18 （CK18）, cytokeratin-19 （CK19） and albumin （ALB） in liver tissues of all mice. Results The expressions of CK18, CK19 and ALB in injured liver tissues were all positive, and the expressions of experimental group B were higher than those of experimental group A （P〈0.05）, but the expressions of CK18, CK19 and ALB in the liver tissues of control group and experimental group C, whose were not injured with CCl4, were all negative. Conclusion Human umbilical cord blood-derived stem cells can differentiate into hepatocytes and express ALB under special microenvironment after liver injured by CCl4 , and the expression level of ALB maybe directly related to the number of human umbilical cord blood stem cells.

关 键 词：脐血 干细胞 肝细胞 分化 

分 类 号：Q25[生物学—细胞生物学]