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

Abstract

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.

The Impact of GFP Reporter Gene Transduction and Expression on Metabolomics of Placental Mesenchymal Stem Cells Determined by UHPLC-Q/TOF-MS

“…Green fluorescent protein (GFP) has been used to evaluate cellular engraftment and for long-term cell tracking [ 3 , 4 ]. Our previous studies showed that transfection of human placental mesenchymal stem cells (hPMSCs) with the GFP gene did not affect their viability, phenotype profile, or pluripotency [ 5 ]. However, the impact of GFP labeling on the metabolism of MSCs remains unknown.…”

Mesenchymal Stem Cells: A Future Option for Intervening Disease Management

“…These properties are not only observed in in-vitro conditions but also in some in-vivo small animal studies, which have revealed the transformation (differentiation) of the MSCs into adult lineages [68]. These are further explained with the presence of the tracker-like GFP [69], indicating the newly formed cells with the presence of the GFP. In human studies, many types of the MSCs expressing pancreatic duodenal homeobox 1 (Pdx1) gene have been shown to differentiate into insulin-producing cells or functions of pancreatic β cells [70].…”

In Vitro Osteogenic Potential of Green Fluorescent Protein Labelled Human Embryonic Stem Cell‐Derived Osteoprogenitors

“…Previous studies have shown successful transduction using lentiviral vectors encoding fluorescent proteins in human adipose-derived stem cells with potential for adipogenic and osteogenic differentiation [32] and in human placenta-derived MSCs with potential for adipogenic, osteogenic, and hepatic differentiation [33]. In a recent study, Ovchinnikov et al used a lentiviral-based delivery system to generated transgenic pluripotent stem cells (PSC) lines that expressed pluripotency-driven GFP [13].…”