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

Abstract

Pancreatic tissue engineering as a therapeutic option for restoring and maintenance of damaged pancreas function has a special focus to using synthetic Scaffolds. This study was designed to evaluate pancreatic differentiation of human induced pluripotent stem cells (hiPSCs) on poly-L-lactic acid and polyvinyl alcohol (PLLA/PVA) scaffolds as 3 D matrix. During differentiation process, morphology of cells gradually changed and iPSCs derived insulin producing cells (iPSCs-IPCs) formed spherical shaped cell aggregation that was the typical shape of islets of pancreas. The highly efficient differentiation of iPSCs into a relatively homogeneous population of IPCs was shown by immunostaining. Real-time reverse transcription polymerase chain reaction (RT-PCR) results demonstrated that iPSCs-IPCs expressed pancreas-specific transcription factors (Pdx1, insulin, glucagon and Ngn3). The expressions of these transcription factors in PLLA/PVA scaffold were significantly higher than 2 D groups. Furthermore, we showed that concentration of insulin and C-peptide in PLLA/PVA scaffold and/or 2 D culture in response to various concentrations of glucose increased but the difference between them were not significant. Altogether the current results demonstrated that PLLA/PVA scaffold could provide the microenvironment that promotes the pancreatic differentiation of iPSCs, up-regulate pancreatic-specific transcription factors and improved metabolic activity.

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

“…Additionally, similar to Lahmy, in this work we demonstrated that endocrine marker of insulin was secreted from induced cells. Briefly, our findings were similar to Enderami [13], who showed that insulin secretion and C-peptide releasing in 3 D medium is higher than 2 D culture in 25 Mm glucose. As well as, our result is opposite to that of Enderami and Abazari (authors) who revealed that pancreatic marker Glut-2 has higher level in 3 D compared to 2 D it can be due to Figure 6.…”

“…In our study we: (1) made polymers that support long-term attachment, proliferation and differentiation of hiPSCs (2) successfully generated the insulin-producing cells from hiPSCs on hybrid scaffold. Similar to Enderami et al [13] our result showed that hiPSCs can tightly stick to scaffolds and penetrate to this hybrid. Also higher expression of pluripotent stem cell marker OCT-4 confirmed the intact characterization of these cells on hybrid scaffold in compared to gelatin coated cells was shown.…”

“…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].…”

Decellularized Wharton's jelly extracellular matrix as a promising scaffold for promoting hepatic differentiation of human induced pluripotent stem cells

“…33 Human iPSCs were cultured and expanded on a feeder layer of mitomycine-C (Sigma-Aldrich) inactivated mouse embryonic fibroblast (MEF) in iPSCs medium (Dulbecco's modified Eagle medium [DMEM]/F12 supplemented with 20% knockout serum replacement, 0.1 mmol/L nonessential amino acids, 2 mM l-glutamine, 0.1 mM β-mercaptoethanol, and 10 ng/mL of recombinant human basic fibroblast growth factor [bFGF], all from Invitrogen, Waltham, MA) in a 5% CO 2 and 95% humidity. 33 Human iPSCs were cultured and expanded on a feeder layer of mitomycine-C (Sigma-Aldrich) inactivated mouse embryonic fibroblast (MEF) in iPSCs medium (Dulbecco's modified Eagle medium [DMEM]/F12 supplemented with 20% knockout serum replacement, 0.1 mmol/L nonessential amino acids, 2 mM l-glutamine, 0.1 mM β-mercaptoethanol, and 10 ng/mL of recombinant human basic fibroblast growth factor [bFGF], all from Invitrogen, Waltham, MA) in a 5% CO 2 and 95% humidity.…”

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

“…Another approach is to use tissue engineering that comprising of three items, including scaffold, cells and growth factors. In this case, tissue engineered constructs could be the content of stem cells‐derived insulin producing cells (IPCs) that grew and differentiated on the polymeric scaffold (Enderami et al, ; Khorsandi, Nejad‐Dehbashi, Ahangarpour, & Hashemitabar, ; Mobarra, Soleimani, Pakzad, Enderami, & Pasalar, ). Among the polymers, those having higher biocompatibility and degradability are preferable.…”