Differentiation of conjunctiva mesenchymal stem cells into secreting islet beta cells on plasma treated electrospun nanofibrous scaffold

Abstract

Transplantation of stem cells using biocompatible nanofibrous scaffolds is a promising therapeutic method for treating Diabetic Mellitus. The aim of this study was to derive insulin-producing cells (IPCs) from conjunctiva-derived mesenchymal stem cell (CJMSCs) and to compare the functionality of differentiated IPCs in a three-dimensional (3D) culture with 2D. Furthermore, the effects of hydrophobicity of scaffold on IPCs differentiation were examined. Scanning electron microscopy (SEM), quantitative real times PCR (qPCR), Immunostaining and flow cytometry were used to analyze fabricated scaffold and the presence of IPCs. Functional maturity of differentiated cells was determined by measuring insulin release and the creation of IPCs was confirmed via gene and protein expression. In this study, the induced CJMSCs were morphologically similar to pancreatic islet-like cells. The expression of the islet-associated genes (glucagon, insulin and Pdx-1) and the insulin release (2.5-fold) in 3D-cultured cells was significantly higher than the 2D. The expression of IPCs genes was significantly higher in CJMSCs differentiated on plasma-treated nanofibers compared to those on untreated scaffolds. In conclusion, the results show that CJMSCs might be a new source for Diabetic Mellitus therapy and the nanofibrous scaffold could be used as a potential cell carrier for islet tissue engineering.

Generation of insulin-producing cells from human induced pluripotent stem cells on PLLA/PVA nanofiber scaffold

“…Previous studies have shown that 3 D cultures increase global gene expression, cell differentiation and cell proliferation. It has been stated that in 3 D culture, these cellular behavior may be related to physiological interactions of cultured cells with neighboring cells and extracellular matrix [28][29][30][31][32]. Besides the increase in the expression of genes in mRNA and protein level which are determinant in differentiation of IPCs and production of insulin in 3 D culture (as shown here), Baharvand et al have shown that the involvement of cellular organelles in protein synthesis also increase [33].…”

Generation of high-yield insulin producing cells from human-induced pluripotent stem cells on polyethersulfone nanofibrous scaffold

“…The result of this study showed expression of these factors was higher in cells cultured on PES nanofibrous scaffold compared to those cultured as two-dimensional in tissue culture plates. Different studies showed that in the PES scaffolds microenvironment, global gene expression, differentiation and proliferation increased [17,[26][27][28]. Furthermore, in this circumstance, cellular behaviour altered due to cellular interactions to neighbouring cells and extracellular matrix [29][30][31].…”

The emerging field of pancreatic tissue engineering: A systematic review and evidence map of scaffold materials and scaffolding techniques for insulin-secreting cells

“…97 The spun fibers offer the possibility of fabricating scaffolds with a high surface to volume ratio, adaptable porosity properties, and applicability for a broad spectrum of materials. 97,98 Researchers used this relatively simple technique to produce polymer fibers from PLA, 99–101 PVA, 53,99,100,102 PCL, 53,96,103,104 polyglecaprone (PGC), 96 PEOT/PBT, 90 gelatin, 105 polyethersulfone, 106,107 silk fibroin, and collagen. One disadvantage of this fabrication method might be the difficulty of scaling up these scaffolds to allow therapeutic human dosages.…”