Enhanced Survival and Function of Islet-Like Clusters Differentiated from Adipose Stem Cells on a Three-Dimensional Natural Polymeric Scaffold: AnIn VitroStudy

Abstract

Autologous adipose stem cells owing to its pluripotent nature offer a valuable source for pancreatic beta cell replacement in the treatment of diabetes mellitus. However, maintaining longevity and functionality of stem cell-derived islet-like cells for long-term in vitro culture is challenging. Signaling interaction between islets and surrounding extracellular matrix (ECM) is an important factor for islet survival and function. Tissue engineering strategy to use scaffolds as substitute for ECM is a key to the problem. In the present study, we fabricated a three-dimensional (3D) biodegradable scaffold comprised of natural polymers dextran and gelatin (DEXGEL) for differentiation of adipose stem cells to islet-like clusters (ILCs). Adipose stem cells derived from subcutaneous fat of New Zealand white rabbits were differentiated to ILCs on DEXGEL scaffold and two-dimensional (2D) culture plates via three stage protocol using cocktail of growth factors. The ILCs differentiated on DEXGEL scaffold exhibited characteristic islet morphology, and expressed islet-specific hormones (insulin, glucagon, and somatostatin). The insulin secretion in response to glucose challenge and viability of ILCs on DEXGEL scaffold were significantly higher in comparison to ILCs on 2D culture. Our results demonstrated for the first time that DEXGEL scaffold simulated an extracellular environment for effective differentiation of rabbit adipose stem cells to ILCs.

Mesenchymal Stem Cells in the Treatment of Type 1 Diabetes Mellitus

“…Moreover, most of these studies failed to generate a sufficient amount of islets for human transplantation and long-term stability. Recent progress in tissue engineering suggested that a biocompatible scaffold may be necessary for the in vitro generation of artificial islets with functional vasculature from stem cells [73]. In our recent study [74], we confirmed that the administration of MSCs into the pancreatic microenvironment can indeed improve the symptoms of diabetes.…”

Engineering of microscale three-dimensional pancreatic islet modelsin vitroand their biomedical applications

“…In another example, adipose stem cells seeded in a 3D biodegradable scaffold composed of natural polymers dextran and gelatin can effectively differentiate to islet-like clusters expressing islet-specific hormones (e.g. insulin, glucagon, and somatostatin) (Aloysious & Nair, 2013). With the scaffold method, the 3D organization of b cells can be controlled and their functions can be improved through cell-cell interaction by tuning the scaffold architecture.…”

Tissue‐engineered islet‐like cell clusters generated from adipose tissue‐derived stem cells on three‐dimensional electrospun scaffolds can reverse diabetes in an experimental rat model and the role of porosity of scaffolds on cluster differentiation

“…Differentiation was accomplished following our published protocol (Aloysious & Nair, 2014). Differentiation was accomplished following our published protocol (Aloysious & Nair, 2014).…”

“…This is highly advantageous for efficient nutrient and oxygen exchange for the cells cultured in these 3D scaffolds (Aloysious & Nair, 2014;Vaikkath et al, 2016). This high water absorption capability can be attributed to the hygroscopic nature of PCL-PTHF-PCL triblock copolymer and wicking action of more number of nanofibers in the lattice scaffold.…”

“…This polymer blend was fabricated in the form of a 3D porous electrospun scaffold by a two-step method so as to overcome the limitation of cell infiltration, allow cell aggregation and formation of 3D clusters of islets after differentiation. The in vitro experiments show that the porous 3D scaffolds promote cell clustering and differentiation of rat adipose-derived mesenchymal stem cells to islet-like clusters with the use of in-house developed differentiation protocol (Aloysious & Nair, 2014). Patterned metallic collectors of varying sizes were used to develop scaffolds of two different pore size ranges.…”