Sahin M Behnan, Schwartz Robert E, Buckley Shannon M, Heremans Yves, Chase Lucas, Hu Wei-Shou, Verfaillie Catherine M
Stem Cell Institute and Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
Liver Transpl. 2008 Mar;14(3):333-45. doi: 10.1002/lt.21380.
Widespread use of liver transplantation in the treatment of hepatic diseases is restricted by the limited availability of donated organs. One potential solution to this problem would be isolation and propagation of liver progenitor cells or stem cells. Here, we report on the isolation of a novel progenitor cell population from unmanipulated (that is, no prior exposure to chemicals and no injury) adult rat liver. Rat liver cells were cultured following a protocol developed in our laboratory to generate a unique progenitor cell population called liver-derived progenitor cells (LDPCs). LDPCs were analyzed by fluorescence-activated cell sorting, real-time polymerase chain reaction (RT-PCR), immunostaining and microarray gene expression. LDPCs were also differentiated into hepatocytes and biliary epithelium in vitro and examined for mature hepatic markers and urea and albumin production. These analyses showed that, LDPCs expressed stem cell markers such as cluster domain (CD)45, CD34, c-kit, and Thy 1, similar to hematopoietic stem cells, as well as endodermal/hepatic markers such as hepatocyte nuclear factor (HNF)3beta, hematopoietically-expressed homeobox gene-1, c-met, and transthyretin. LDPCs were negative for OV-6, cytokeratins (CKs), albumin, and HNF1alpha. The microarray gene expression profile demonstrated that they showed some similarities to known liver progenitor/stem cells such as oval cells. In addition, LDPCs differentiated into functional hepatocytes in vitro as shown by albumin expression and urea production. In conclusion, LDPCs are a population of unique liver progenitors that can be generated from unmanipulated adult liver, which makes them potentially useful for clinical applications, especially for cell transplantation in the treatment of liver diseases.
肝脏移植在治疗肝脏疾病方面的广泛应用受到捐赠器官有限可用性的限制。解决这一问题的一个潜在办法是分离和增殖肝祖细胞或干细胞。在此，我们报告从未经处理（即未事先接触化学物质且未受伤）的成年大鼠肝脏中分离出一种新型祖细胞群体。按照我们实验室制定的方案培养大鼠肝细胞，以产生一种称为肝源性祖细胞（LDPCs）的独特祖细胞群体。通过荧光激活细胞分选、实时聚合酶链反应（RT-PCR）、免疫染色和微阵列基因表达对LDPCs进行分析。LDPCs在体外也分化为肝细胞和胆管上皮细胞，并检测其成熟肝脏标志物以及尿素和白蛋白的产生。这些分析表明，LDPCs表达干细胞标志物，如与造血干细胞相似的集落结构域（CD）45、CD34、c-kit和Thy 1，以及内胚层/肝脏标志物，如肝细胞核因子（HNF）3β、造血表达同源盒基因-1、c-met和转甲状腺素蛋白。LDPCs对OV-6、细胞角蛋白（CKs）、白蛋白和HNF1α呈阴性。微阵列基因表达谱表明，它们与已知的肝祖细胞/干细胞如卵圆细胞有一些相似之处。此外，如白蛋白表达和尿素产生所示，LDPCs在体外分化为功能性肝细胞。总之，LDPCs是一群独特的肝祖细胞，可从未经处理的成年肝脏中产生，这使其在临床应用中具有潜在用途，特别是在治疗肝脏疾病的细胞移植方面。
