Lee Sum P, Savard Christopher E, Kuver Rahul
Division of Gastroenterology, Department of Medicine, University of Washington, Puget Sound Veterans Affairs Health Care System, Seattle, WA 98195, USA.
Am J Pathol. 2009 Mar;174(3):842-53. doi: 10.2353/ajpath.2009.080262. Epub 2009 Feb 13.
We tested the hypothesis that well-differentiated gallbladder epithelial cells (GBECs) are capable of engrafting and surviving in murine liver and acquire phenotypic characteristics of hepatocytes. GBECs isolated from transgenic mice that constitutively express green fluorescent protein (GFP) were either cultured before transplantation or transplanted immediately following isolation. Recipient mice with severe-combined immunodeficiency underwent retrorsine treatment and either partial hepatectomy before transplantation or carbon tetrachloride treatment following transplantation. From 1 to 4 months following transplantation, the livers of recipient mice contained discrete colonies of GFP(+) cells. Most GFP(+) cells surrounded vesicles, were epithelial cell-like in morphology, and expressed the biliary epithelial markers cytokeratin 19 and carbonic anhydrase IV. Subpopulations of GFP(+) cells resembled hepatocytes morphologically and expressed the hepatocyte-specific markers connexin-32 and hepatic nuclear factor-4alpha, but not cytokeratin 19 or carbonic anhydrase IV. At 4 months, cells in GFP(+) colonies were not actively proliferating as determined by proliferating cell nuclear antigen expression. Thus, GBECs are capable of engrafting and surviving in damaged mouse livers, and some can differentiate into cells with hepatocyte-like features. These findings suggest that environmental cues in the recipient liver are sufficient to allow a subpopulation of donor GBECs to differentiate into hepatocyte-like cells in the absence of exogenous transcriptional reprogramming. GBECs might be used as donor cells in a cell transplantation approach for the treatment of liver disease.
高分化的胆囊上皮细胞(GBECs)能够在小鼠肝脏中植入并存活,并获得肝细胞的表型特征。从组成型表达绿色荧光蛋白(GFP)的转基因小鼠中分离出的GBECs,要么在移植前进行培养,要么在分离后立即移植。患有严重联合免疫缺陷的受体小鼠接受了倒千里光碱治疗,并在移植前进行了部分肝切除术,或在移植后进行了四氯化碳处理。移植后1至4个月,受体小鼠的肝脏中含有离散的GFP(+)细胞集落。大多数GFP(+)细胞围绕着囊泡,形态上类似上皮细胞,并表达胆管上皮标志物细胞角蛋白19和碳酸酐酶IV。GFP(+)细胞亚群在形态上类似于肝细胞,并表达肝细胞特异性标志物连接蛋白-32和肝细胞核因子-4α,但不表达细胞角蛋白19或碳酸酐酶IV。在4个月时,通过增殖细胞核抗原表达确定,GFP(+)集落中的细胞未积极增殖。因此,GBECs能够在受损的小鼠肝脏中植入并存活,并且一些细胞可以分化为具有肝细胞样特征的细胞。这些发现表明,受体肝脏中的环境信号足以使供体GBECs亚群在没有外源性转录重编程的情况下分化为肝细胞样细胞。GBECs可能用作细胞移植方法中的供体细胞来治疗肝脏疾病。