Differentiation of PDX1 gene-modified human umbilical cord mesenchymal stem cells into insulin-producing cells in vitro

Abstract

Abstract. Mesenchymal stem cells (MSCs) have significant advantages over other stem cell types, and greater potential for immediate clinical application. MSCs would be an interesting cellular source for treatment of type 1 diabetes. In this study, MSCs from human umbilical cord were differentiated into functional insulin-producing cells in vitro by introduction of the pancreatic and duodenal homeobox factor 1 (PDX1) and in the presence of induction factors. The expressions of cell surface antigens were detected by flow cytometry. After induction in an adipogenic medium or an osteogenic medium, the cells were observed by Oil Red O staining and alkaline phosphatase staining. Recombinant adenovirus carrying the PDX1 gene was constructed and MSCs were infected by the recombinant adenovirus, then treated with several inducing factors for differentiation into islet β-like cells. The expression of the genes and protein related to islet β-cells was detected by immunocytochemistry, RT-PCR and Western blot analysis. Insulin and C-peptide secretion were assayed. Our results show that the morphology and immunophenotype of MSCs from human umbilical cord were similar to those present in human bone marrow. The MSCs could be induced to differentiate into osteocytes and adipocytes. After induction by recombined adenovirus vector with induction factors, MSCs were aggregated and presented islet-like bodies. Dithizone staining of these cells was positive. The genes' expression related to islet β-cells was found. After induction, insulin and C-peptide secretion in the supernatant were significantly increased. In conclusion, our results demonstrated that PDX1 gene-modified human umbilical cord mesenchymal stem cells could be differentiated into insulin-producing cells in vitro.

The role of DNA methylation in thermogenic adipose biology

“…On the other hand, brown adipocytes have hypomethylated exonic regions that are significantly enriched for genes involved in brown fat functions such as the mitochondrial respiratory chain and fatty acid oxidation [25]. Notably, several Hox transcription factors are differentially methylated, some of which are linked to adipogenesis and diabetes [26,27]. For example, Hoxc9 is a well-established adipocyte marker [28], and Hoxc9 and Hoxc10 promoter methylation is inversely correlated with their gene expression in brown adipose tissue.…”

The progress of Stem cells in the treatment of diabetes mellitus type 1

“…They also showed that β-cell related genes were expressed in both differentiated cells and β-like cells transplanted in to the liver of STZ-induced diabetic rats through portal vein. As a result blood glucose levels were significantly reduced 4weeks after transplantation (He et al, 2011). The differentiated IPCs from human WJ-MSCs could alleviate hyperglycemia in diabetic mice (Tsai et al, 2012).…”

“…Expression of insulin Secretion of C-peptide Expression of pancreas-specific genes (Hashemian et al,2015) Correspondence to high concentrations of glucose (Wu et al, 2009) Reduction of blood glucose levels after 4weeks of transplantation (He et al,2011) (Hashemian et al, 2015). …”

Defined three-dimensional culture conditions mediate efficient induction of definitive endoderm lineage from human umbilical cord Wharton’s jelly mesenchymal stem cells

“…Another group managed to differentiate WJ-MSCs into insulin-producing cells in vitro [12]. Other groups reported the differentiation of WJ-MSCs into insulin-producing cells with islet-like morphology, using recombinant adenovirus– PDX1 gene constructs [13, 14]. Despite showing positive indications toward DE differentiation, these studies reported the use of animal serum and/or genetic modifications, and resulted in low differentiation capacities.…”