Human umbilical cord blood-derived cells differentiate into hepatocyte-like cells in the Fas-mediated liver injury model.
Human umbilical cord blood-derived cells differentiate into hepatocyte-like cells in the Fas-mediated liver injury model.
复制标题
DOI：
10.1152/ajpgi.00049.2005
复制
发表时间：
2005-12
期刊：
影响因子：
--
通讯作者：
K. Nonome;Xiao‐Kang Li;T. Takahara;Y. Kitazawa;N. Funeshima;Y. Yata;F. Xue;Masami Kanayama;E. Shinno;Chieko Kuwae;S. Saito;A. Watanabe;T. Sugiyama
中科院分区：
文献类型：
--
作者：
K. Nonome;Xiao‐Kang Li;T. Takahara;Y. Kitazawa;N. Funeshima;Y. Yata;F. Xue;Masami Kanayama;E. Shinno;Chieko Kuwae;S. Saito;A. Watanabe;T. Sugiyama
人脐血(HUCB)含有干细胞/祖细胞，可分化为多种细胞类型。在这项研究中，我们研究了HUCB细胞在体外和体内是否分化为肝细胞。我们还检测了CD34是否可以作为干细胞对肝细胞的选择标记。取正常足月新生儿脐血，分离CD34(+/-)细胞。体外培养HUCB细胞4wk，检测肝组织特异性基因的表达。在体内研究中，非肥胖的糖尿病/严重联合免疫缺陷小鼠通过Fas配体携带的腺病毒载体造成肝损伤或仅进行辐射。小鼠同时接受或不接受HUCB、CD34(+)或CD34(-)细胞移植。4wk后，检测人特异性基因/蛋白的表达。在体外研究中，人类肝脏特异性基因在培养7天后呈阳性。免疫荧光研究显示，甲胎蛋白、细胞角蛋白19和白蛋白在圆形细胞中呈阳性染色。在体内研究中，免疫组织化学分析显示Fas配体/移植组小鼠肝脏中有人白蛋白阳性、肝细胞特异性抗原阳性细胞。使用人类Y染色体的荧光原位杂交分析也显示出阳性信号。然而，移植细胞类型之间没有发现差异。相反，照射/移植组未检测到免疫阳性细胞。RT-PCR结果还显示，只有Fas配体/移植组有人肝细胞特异性基因的表达。HUCB细胞在小鼠肝脏中分化为肝细胞样细胞，在此过程中肝损伤是必不可少的。CD34(+)和CD34(-)细胞在人肝细胞特异性表达上没有差异。
Human umbilical cord blood (HUCB) contains stem/progenitor cells, which can differentiate into a variety of cell types. In this study, we investigated whether HUCB cells differentiate into hepatocytes in vitro and in vivo. We also examined whether CD34 could be the selection marker of stem cells for hepatocytes. HUCB cells were obtained from normal full-term deliveries, and CD34(+/-) cells were further separated. For in vitro study, HUCB cells were cultured for 4 wk, and expressions of liver-specific genes were examined. For the in vivo study, nonobese diabetic/severe combined immunodeficient mice were subjected to liver injury by a Fas ligand-carried adenoviral vector or only radiated. Mice were treated simultaneously with or without cell transplantation of HUCB, CD34(+), or CD34(-) cells. After 4 wk, human-specific gene/protein expression was examined. In the in vitro study, human liver-specific genes were positive after 7 days of culture. The immunofluorescent study showed positive staining of alpha-fetoprotein, cytokeratin 19, and albumin in round-shaped cells. In the in vivo study, immunohistochemical analysis showed human albumin-positive, hepatocyte-specific antigen-positive cells in mouse livers of the Fas ligand/transplantation group. Fluorescence in situ hybridization analysis using the human Y chromosome also showed positive signals. However, no difference between transplanted cell types was detected. In contrast, immunopositive cells were not detected in the irradiated/transplantation group. The RT-PCR result also showed human hepatocyte-specific gene expressions only in the Fas ligand/transplantation group. HUCB cells differentiated into hepatocyte-like cells in the mouse liver, and liver injury was essential during this process. The differences between CD34(+) and CD34(-) cells were not observed in human hepatocyte-specific expression.