Improvement of mesenchymal stem cell differentiation into the endoderm lineage by four step sequential method in biocompatible biomaterial

Abstract

Introduction: The goal of the study described here, was to investigate the potential of umbilical cord derived mesenchymal stem cell (UC-MSCs) into hepatocyte like cells in a sequential 2D and 3D differentiation protocols as alternative therapy. Methods: Mesenchymal stem cells (MSCs) were isolated from the umbilical cord (UC) and CD markers were analyzed by flow cytometry. For hepatic differentiation of UC-MSCs, cells were induced with a sequential 4-step protocol in 3D and 2D culture system. Urea concentration and albumin secretion into the culture medium was quantified by ELISA. Gene expression levels of AFP, ALB, and CK18 were determined by RT-PCR. Data were statistically analyzed by the SPSS software. The difference between the mean was considered significant when p < 0.05. Results: Growth factor dependent morphological changes from elongated fibroblast-like cells to round epithelial cell morphology were observed in 2D culture. Cell proliferation analysis showed round-shaped morphology with clear cytoplasm and nucleus on the alginate scaffold in 3D culture. The mean valuses of albumin production and urea secretion were significantly higher in the 3D Culture system when compared with the 2D culture (p = 0.005 vs p = 0.001), respectively. Treatment of cells with TSA in the final step of differentiation induced an increased expression of CK18 and a decreased expression of αFP in both the 3D and 2D cultures (p = 0.026), but led to a decreased albumin gene expression, and an increased expression in the 2D culture (p = 0.001). Conclusion: Findings of the present study indicated that sequential exposure of UC-MSCs with growth factors in 3D culture improves hepatic differentiation.

Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells Are Functionally and Genetically Different From Bone Marrow-Derived Mesenchymal Stromal Cells

“…Conversely, the high expression of both CDH1 and SOX7 in BM-MSCs (Fig. 5) may infer the existence of an endodermal precursor which is in line with the potential of these cells to transdifferentiate into endodermal lineages [50]. Overall, a similarly low level of MIXL1 detected on both cell types implies their…”

Inhibition of the RhoGTPase Cdc42 by ML141 enhances hepatocyte differentiation from human adipose-derived mesenchymal stem cells via the Wnt5a/PI3K/miR-122 pathway: impact of the age of the donor

“…Other studies reported that TSA may reverse the age impact within other human MSC subtypes and inhibits in-vitro differentiation of BMSCs into neuronal lineage cells [ 73 , 74 , 81 ]. Cdc42 inhibition was described to induce hepatic differentiation and maturation in UCMSCs involving CK-18 and other key hepatic genes [ 82 , 83 ].…”

Current progress in hepatic tissue regeneration by tissue engineering

“…It was found that these spheroids can partially support liver function in hepatectomized animal model after subcutaneous or intraperitoneal transplantation [34]. Similar to iPSCs and ESCs, MSCs also have shown a high hepatic differentiation potential either in vivo or in vitro model [35]. Bone marrow-derived MSCs demonstrated an enhanced expression of hepatocyte-specific markers and exhibited hepatocellular function after introduction to the liver decellularized scaffold in the presence of EGF and HGF ( Fig.…”