In-vitro generation of human adipose tissue derived insulin secreting cells: up-regulation of Pax-6, Ipf-1 and Isl-1

Abstract

We present a study of up-regulation of genes responsible for pancreatic development in glucose-sensitive insulin-secreting mesenchymal stem cells (IS-MSC) generated and differentiated from human adipose tissue (h-AD), with use of our specific differentiation media and without use of any xenogenic material. Anterior wall abdominal fat was collected from 56 volunteers and cultured in selfdesigned proliferation medium for 10 days. Cells were harvested by trypsinization and differentiated into insulin-expressing cells using self-designed differentiation medium for 3 days followed by evaluation for transcriptional factors Pax-6, Ipf-1, Isl-1, C-peptide and insulin secretion. Generated IS-MSC showed expression of Pax-6, Pdx-6 and Isl-1. Non-differentiated MSC as well as their further culture in absence of differentiation medium were used as negative controls. Generated 56 IS-MSC cell-lines were glucose responsive i.e. mean C-Peptide and insulin secretion levels were measured 0.41 ng/ml and 13.13 lU/ml, respectively, in absence of glucose which rose to 1.18 ng/ml and 83.42 lU/ml, respectively, following glucose challenge (p \ 0.001). The mean rise in C-peptide and insulin secretion levels was 2.88 and 6.35 fold, respectively. To conclude insulin-secreting h-AD-MSC can be generated safely and effectively with application of specific differentiation media without xenogeneic material/any genetic modification, showing expression of transcriptional factors Pax-6, Ipf-1 and Isl-1.

Immunomodulatory Effect of Adipose-Derived Stem Cells: The Cutting Edge of Clinical Application

“…Adipose-derived mesenchymal stem cells (ASCs) represent a population of self-renewing multipotent adult cells in the vascular stroma of adipose tissues, playing important roles in development, post-natal growth, maintenance of tissue homeostasis and tissue repair and regeneration (Shingyochi et al, 2015;Dai et al, 2016). Significant amount of ASCs can be readily accessible from subcutaneous liposuction and, when appropriately stimulated, they can further differentiate into several cell-like types, including adipocytes, osteocytes, neural cells, vascular endothelial cells, cardiomyocytes, pancreatic cells, and hepatocytes (Zuk et al, 2001(Zuk et al, , 2002Planat-Bénard et al, 2004;Aurich et al, 2009;Dave et al, 2014;Sommese et al, 2017). To date, mechanisms responsible for ASC therapeutic efficacy have been only partially investigated.…”

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

“…Moreover, they can differentiate into specific cells such as fat cells, osteocytes, chondrocytes and myocytes. Importantly, these cells of mesodermal origin can be driven to differentiate into endodermal cells such as insulin-producing cells via trans-differentiation (Chandra et al, 2009;Dave et al, 2013Dave et al, , 2014Fedyunina et al, 2011;Hwang et al, 2011;Marappagounder et al, 2013;Mohamad Buang et al, 2012;Moshtagh et al, 2013;Timper et al, 2006). This study aimed to develop and test an efficient protocol to produce functional islets that can be used in research and application.…”

Obesity and Diabetes

“…Cells were administered intra-portally. In vitro generated ISC showed presence of pancreatic transcriptional factors Pax-6, Isl-1, and Pdx-1(by immunoflurescence assay) (Dave et al 2014). Chemiluminescence assay detected secretion of glucose and C-peptide in response to glucose concentration in vitro.…”