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

Abstract

Transplantation of islet is a promising method in treatment of patients with type 1 diabetes mellitus (T1DM), however, is limited by islet shortage. The aim of this study was to prepare a polyethersulfone (PES) nanofibrous scaffolds to evaluate the pancreatic differentiation of human induced pluripotent stem cells (hiPSCs). The differentiation process in tissue culture dishes and PES scaffolds was evaluated at mRNA and protein level by RT-qPCR and immunofluorescence assay, respectively. The functionality of differentiated cells was determined by insulin and C-peptide release in response to glucose challenges. The results of this study showed that cells cultured on PES nanofibrous scaffolds exhibit more pancreatic b-cell characteristics as they express more pancreatic tissue-specific genes and proteins. Furthermore, the immunoassay showed that differentiated cells in both culture plates and PES scaffolds groups are functional and secrete C-peptide and insulin in response to glucose challenges. Altogether, the results of this study demonstrated that PES nanofibrous scaffold could provide the microenvironment that promotes the differentiation of induced pluripotent stem cells (iPSCs) into insulin producing cells.

Comparison of human‐induced pluripotent stem cells and mesenchymal stem cell differentiation potential to insulin producing cells in 2D and 3D culture systems in vitro

“…The iPSCs have also been introduced for use in cell therapy and regenerative medicine in the last decade, and their ability to differentiate into many other cells, including IPCs, has been reported previously (Mansour et al, 2018;Morizane et al, 2015;Song, Wang, Li, & Yang, 2016;Yamanaka, 2012).…”

Three-dimensional nanofiberous PLLA/PCL scaffold improved biochemical and molecular markers hiPS cell-derived insulin-producing islet-like cells

“…But our result showed that pancreatic markers are markedly expressed well in PLLA/PCL hybrid scaffold than they reported. Poly (e-caprolactone) (PCL) is a bioresorbable, biocompatible and inexpensive nanofiber, having native tissue like properties that are generally used for tissue regeneration, as a reported, because of low bioactivity, use of pure PCL in tissue engineering is limited [7,15,16] . Zhang et al, reported that in order to enhance adhesion, proliferation and encourage of cell to growth into the PCL scaffolds, this nanofibres need effective hybridization with other bioactive molecules compared to single PCL [32].…”

“…However, problems such as donor organ shortage, high risks of surgery and Immunologic transplant rejection have limited the successful development of this treatment. Thus, the novel alternative strategies such as gene transfer [7] and usage of induced pluripotent stem cell, as well as three-dimensional (3D) culture of nanofibrous scaffolds should be developed to overcome these limitations [8]. Nevertheless, the first method is expensive, difficult and time consuming.…”

“…This condition makes them more practical than two-dimensional culture [12]. Recently, a number of studies have used various scaffolds such as PES [7], PLLA/PVA [13] and PCL/PVA [14] to obtain human iPSCs insulin-producing beta cells. However, some of the applied scaffolds such as PES have limitations, like being a non-biodegradable polymer it requested for repairing hard damaged tissues like bone and cartilage [9,15].…”

“…However, some of the applied scaffolds such as PES have limitations, like being a non-biodegradable polymer it requested for repairing hard damaged tissues like bone and cartilage [9,15]. Moreover, PES has a low potential to attract cells to penetrate into the nanofibrous structure [7]. Another used scaffold is PCL; this nanofibrous is highly hydrophobic thus resulting in poor cell attachment.…”