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GFP Labeling and Hepatic Differentiation Potential of Human Placenta-Derived Mesenchymal Stem Cells

机译：人胎盘来源的间充质干细胞的GFP标记和肝分化潜能

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Background: Stem cell-based therapy in liver diseases has received increasing interest over the past decade, but direct evidence of the homing and implantation of transplanted cells is conflicting. Reliable labeling and tracking techniques are essential but lacking. The purpose of this study was to establish human placenta-derived mesenchymal stem cells (hPMSCs) expressing green fluorescent protein (GFP) and to assay their hepatic functional differentiation in vitro. Methods: The GFP gene was transduced into hPMSCs using a lentivirus to establish GFP+ hPMSCs. GFP+ hPMSCs were analyzed for their phenotypic profile, viability and adipogenic, osteogenic and hepatic differentiation. The derived GFP+ hepatocyte-like cells were evaluated for their metabolic, synthetic and secretory functions, respectively. Results: GFP+ hPMSCs expressed high levels of HLA I, CD13, CD105, CD73, CD90, CD44 and CD29, but were negative for HLA II, CD45, CD31, CD34, CD133, CD271 and CD79. They possessed adipogenic, osteogenic and hepatic differentiation potential. Hepatocyte-like cells derived from GFP+ hPMSCs showed typical hepatic phenotypes. Conclusions: GFP gene transduction has no adverse influences on the cellular or biochemical properties of hPMSCs or markers. GFP gene transduction using lentiviral vectors is a reliable labeling and tracking method. GFP+ hPMSCs can therefore serve as a tool to investigate the mechanisms of MSC-based therapy, including hepatic disease therapy.
机译：背景：在过去的十年中，以干细胞为基础的肝病治疗方法受到越来越多的关注，但是直接证据表明移植细胞的归巢和植入存在矛盾。可靠的标签和跟踪技术是必不可少的，但却缺乏。这项研究的目的是建立表达绿色荧光蛋白（GFP）的人胎盘来源的间充质干细胞（hPMSC），并在体外测定其肝功能分化。方法：利用慢病毒将GFP基因导入hPMSCs，建立GFP + hPMSCs。分析了GFP + hPMSCs的表型特征，生存力以及成脂，成骨和肝细胞分化。分别评估衍生的GFP + 肝细胞样细胞的代谢，合成和分泌功能。结果：GFP + hPMSCs表达高水平的HLA I，CD13，CD105，CD73，CD90，CD44和CD29，但对HLA II，CD45，CD31，CD34，CD133，CD271和CD79呈阴性。它们具有成脂，成骨和肝分化的潜力。 GFP + hPMSCs衍生的肝细胞样细胞表现出典型的肝表型。结论：GFP基因转导对hPMSCs或标志物的细胞或生化特性无不利影响。使用慢病毒载体的GFP基因转导是一种可靠的标记和跟踪方法。因此，GFP + hPMSCs可作为研究基于MSC的治疗机制（包括肝病治疗）的工具。

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