Comparison of insulin secretion by transduced adipose‐derived and endometrial‐derived stem cells in 2D and 3D cultures on fibrin scaffold

Abstract

Type 1 diabetes is a metabolic disorder caused by the loss or dysfunction of β‐cells in the pancreas. Organ shortage is a critical concern of diabetic patients in need of beta islet transplantation. Tissue engineered islets are promising alternatives to traditional organ transplantation. Recent progress in stem cell biology and gene cloning techniques has raised hopes for the generation of insulin producing cells (IPCs) without the need of immunosuppression. The purpose of this study was to produce IPCs using human adipose‐derived stem cells (hADSCs) and human endometrial‐derived stem cells (hEnSCs) and also to compare the level of insulin secretion by these cells in 2D and 3D culture systems on fibrin scaffolding. Stem cells differentiation was carried out through transduction with an insulin over expression lentiviral vector. Real‐time PCR and immunocytochemistry confirmed the successful transduction of both cell types. Both cell types showed comparable insulin secretion by ELISA.3D culture resulted in higher amounts of insulin secretion of the two cell types versus 2D as control. This study showed that insulin gene delivery to the stem cells could be an efficient method for producing IPCs and fibrin encapsulation enhances the functionality of these cells.

A Temperature‐Based Easy‐Separable (TempEasy) 3D Hydrogel Coculture System

“…Note that we also quantified expression of other genes that are introduced later in this work (Figure S1, Supporting Information); in line with FPS1 and CDKN1A, they do not show significant differences in expression levels between the different culture conditions. We highlight that in our matrices, cell behavior does depend on culture dimensionality as was previously found, [65][66][67] but that in terms of gene expression, the different cells show a largely similar response irrespective of their 2D or 3D environment.…”