Short-Term Protocols to Obtain Insulin-Producing Cells from Rat Adipose Tissue: Signaling Pathways and In Vivo Effect

Abstract

Studies using mesenchymal stromal cells (MSCs) as a source of insulin-secreting cells (IPCs) are a promising path in the pursuit for diabetes therapy. Here, we investigate three short-term differentiation protocols in order to generate IPCs from autologous adipose-derived stromal cells (ADSCs) with an expressive insulin-secreting profile in vitro and in vivo, as well as the signaling pathways involved in the chosen differentiation protocols. We extracted and cultured ADSCs and differentiated them into IPCs, using three different protocols with different inductors. Afterwards, the secretory profile was analyzed and IPCs differentiated in exendin-4/activin A medium, which presented the best secretory profile, was implanted in the kidney subcapsular region of diabetic rats. All protocols induced the differentiation, but media supplemented with exendin-4/activin A or resveratrol induced the expression and secretion of insulin more efficiently, and only the exendin-4/activin-A-supplemented medium generated an insulin secretion profile more like β-cells, in response to glucose. The PI3K/Akt pathway seems to play a negative role in IPC differentiation; however, the differentiation of ADSCs with exendin-4/activin A positively modulated the p38/MAPK pathway. Resveratrol medium activated the Jak/STAT3 pathway and generated IPCs apparently less sensitive to insulin and insulin-like receptors. Finally, the implant of IPCs with the best secretory behavior caused a decrease in hyperglycemia after one-week implantation in diabetic rats. Our data provide further information regarding the generation of IPCs from ADSCs and strengthen evidence to support the use of MSCs in regenerative medicine, specially the use of exendin-4/activin A to produce rapid and effectively IPCs with significant in vivo effects.

Mesenchymal Stem Cell Therapy in Diabetic Cardiomyopathy

“…MSCs can differentiate into insulin-producing cells (IPCs) both in vitro and in vivo, which underlies their potential for treating DM [ 98 ]. The transplantation of IPCs derived from different types of MSCs in vitro is effective in regulating blood glucose levels in experimental diabetic animals [ 99 , 100 , 101 ]. However, although MSCs migrate to the pancreas, the differentiation into IPCs is minimal, and it is not the explanation for the possible regenerative effects induced by cell therapy.…”

A One-Step, Monolayer Culture and Chemical-Based Approach to Generate Insulin-Producing Cells From Human Adipose-Derived Stem Cells to Mitigate Hyperglycemia in STZ-Induced Diabetic Rats

“…One study reported that use of a low-attachment suspension culture method could generate IPC spheroids in the presence of activin A and exendin-4 28 . Additionally, Wartchow et al compared three different formulations of differentiationrelated factors and found that IPCs generated by Timper's protocol had the best insulin secretion profile and could ameliorate hyperglycemia in streptozotocin (STZ)-induced diabetic rats 29 , indicating that IPCs generated via the activin A/ exendin-4 formulation have the potential to treat diabetes. However, spheroids that generate spontaneously during culture on low-attachment plates have some disadvantages such as variability in spheroid size and a lack of nutrients and oxygen in the spheroid core, resulting in a skewed distribution of proliferating, non-proliferating, and apoptotic cells that can lead to cell death during culture or after transplantation 30,31 .…”

In-Vitro Differentiation Adipose-Derived Mesenchymal Stem Cells into Pancreatic Progenitor Cells

“…Among the various sources of adult stem cells, adipose tissue is an ideal population of stem cells. Adipose tissue contains an abundance of stem cells, which can be obtained by a procedure that is relatively easy, inexpensive and minimally invasive 5,6 . Several studies were conducted to differentiate MSCs into cells that were similar to pancreatic islets and which, when transplanted, proved to be functional and producing insulin in diabetic rats 6 .…”

“…Adipose tissue contains an abundance of stem cells, which can be obtained by a procedure that is relatively easy, inexpensive and minimally invasive 5,6 . Several studies were conducted to differentiate MSCs into cells that were similar to pancreatic islets and which, when transplanted, proved to be functional and producing insulin in diabetic rats 6 . Pancreatic progenitor cells (PPCs) give rise to all cell types in the pancreas during development 7 .…”

“…Adipose-derived mesenchymal stem cells (ADMSCs) are mesenchymal stem cells (MSCs) which obtained from adipose tissue and had unique characteristics making them suitable for differentiation into endocrine pancreatic progenitor 10,11 . Differentiation MSCs into endocrine progenitor pancreas can be carried out using many extrinsic factors such as nicotinamide, exendin-4, and GLP-1 receptor agonist 6 .…”