Production of islet-like insulin-producing cell clusters in vitro from adiposederived stem cells

Abstract

Abstract-Diabetes mellitus is a high incidence disease that has increased rapidly in recent years. Many new therapies are being studied and developed in order to find an effective treatment. An ideal candidate is stem cell therapy. In this study, we investigated the differentiation of adipose derived stem cells (ADSCs) into pseudo-islets in defined medium in vitro, to produce large quantities of insulin-producing cells (IPCs) for transplantation. ADSCs isolated from adipose tissue were induced to differentiate into islet-like insulin-producing cell clusters in vitro by inducing medium DMEM/F12 containing nicotinamide, N2, B27, bFGF, and insulin-transferrin-selenite (ITS). Differentiated cells were analyzed for properties of IPCs, including storage of Zn 2+ by dithizone staining, insulin production by ELISA and immunochemistry, and beta cell-related gene expression by reverse transcriptase PCR. The results showed that after 2 weeks of differentiation, the ADSCs aggregated into cell clusters, and after 4 weeks they formed islets, 50-400 micrometers in diameter. These islet cells exhibited characteristics of pancreatic beta cells as they were positive for dithizone staining, expressed insulin in vitro and C-peptide in the cytoplasm, and expressed pancreatic beta cellspecific genes, including Pdx-1, NeuroD, and Ngn3. These results demonstrate that ADSCs can be used to produce a large number of functional islets for research as well as application.

In vitro Differentiation of Adipose-Derived Mesenchymal Stem Cell into Insulin-Producing Cells

“…Cells clusters were formed within 15 days of differentiation, in contrary to the study of Karaoz et al, it took them 38 days to induce the formation of clusters after co-culture with isolated islets [19]. The cells were positive DTZ and showed immuno-reactivity to anti-human insulin antibody and responded to high glucose concentration after 15 days of differentiation, consistent with other studies [20], while in Johnson et al, experiments ADSC differentiated into insulin producing cells and DTZ positive clusters were observed after 30 days [21].…”

Mesenchymal Stem Cell Therapy for Diabetic Kidney Disease: A Review of the Studies Using Syngeneic, Autologous, Allogeneic, and Xenogeneic Cells

“…Both diabetic and nondiabetic AT-MSCs were able to differentiate into adipocytes and chondrocytes, yet not exhibiting islet-like cell differentiation in that study [ 132 ]. Importantly, in vitro studies documented the differentiation potential of human AT-MSCs into islet-like cells when these cells were obtained from healthy individuals who underwent abdominoplasty or liposuction [ 133 – 135 ]. Transdifferentiated cells exhibit positive staining for dithizone, increased expression of islet cell-related genes (Pdx-1, Isl1, Ngn3, NeuroD1, Pax4, and GLUT2), and insulin secretion when these cells were challenged with high concentrations of glucose.…”

Modern pancreatic islet encapsulation technologies for the treatment of type 1 diabetes

“…Мезенхимные стволовые клетки. Использование МСК при СД возможно двумя способами: дифференцировкой в инсулинпродуцирующие клетки [88] и прямым введением недифференцированных МСК [139]. МСК жировой ткани при культивировании в средах, содержащих фактор роста фибробластов, способны экспрессировать маркер Isl1, необходимый для образования островковых клеток ПЖ [89].…”