Porcine pancreas mesenchymal cell characterization and functional differentiation into insulin‑producing cells in vitro

Abstract

Cell therapy is a promising treatment strategy for patients with type 1 diabetes. Porcine pancreas-derived mesenchymal stromal cells (PMSCs) have emerged as one of the most widely used cell resources owing to their high proliferative capacity and multi-lineage differentiation potential. Although the induction efficiency and insulin production of induced insulin-producing cells (IPCs) derived from PMSCs have been estimated, these have primarily focused on the function of induced cells and alterations in related gene expression levels. However, morphological analyses and biological characterization of PMSCs and induced IPCs have not been conducted. Therefore, the present study aimed to optimize an induction protocol, resulting in a 78.92% induction rate. The present study investigated the biological characteristics of PMSCs and optimized a simple but functional three-step protocol to transform PMSCs into IPCs. PMSCs were isolated from 2-3-month-old Bama miniature pig embryos, which were then subcultured to passage 16. The surface markers pancreatic and duodenal homeobox 1, NK6 homeobox 1, Vimentin, Nestin, CD73, CD90, neurogenin 3, CD45 and CD34 were detected by immunofluorescence staining or flow cytometry. Proliferative capacity was evaluated by constructing growth curves of cells at three different passages. Functional differentiation was assessed by morphological observation, dithizone staining, and immunofluorescence staining of C-peptide, insulin, NK6 homeobox 1 and glucagon. The production of insulin by differentiated cells was also analyzed by performing ELISAs. The results demonstrated that differentiated cells were distributed with an islet-like structure, expressed specific markers C-peptide and insulin, and displayed glucose responsiveness. The results of the present study demonstrated that PMSCs were functionally induced into IPCs with the optimized three-step protocol, which may serve as a potential cell therapy strategy to widen the availability and promote the clinical application of cell therapy.

Mn(II)-Mediated Self-Assembly of Tea Polysaccharide Nanoparticles and Their Functional Role in Mice with Type 2 Diabetes

“…Immunofluorescence Analysis of Insulin in the Pancreatic β-Cell of T2DM Mice. Since the previous study reported that insulin is secreted by β-cells in the pancreas, which is one of the most important metabolic hormones to control blood glucose, 38,39 we investigated the ability of insulin secretion and the potential effects of MTNP on pancreatic βcell injury in diabetic mice. Immunofluorescence (IF) analysis was utilized to measure the expression of insulin in the pancreas, in which the anti-insulin antibody was used to determine the insulin expression.…”

In Vivo Assessment of the Effects of Mono-Carrier Encapsulated Fucoxanthin Nanoparticles on Type 2 Diabetic C57 Mice and Their Oxidative Stress

“…Insulin is known to be secreted by β-cells in the pancreas and is one of the most important metabolic hormones in the control of blood glucose [ 41 ]. The expression of insulin in the pancreas can be measured using immunofluorescence (ICC) labeling of the insulin in the pancreas (pink fluorescent area in Figure 5 a).…”

Therapeutic Effects of Insulin-Producing Human Umbilical Cord-Derived Mesenchymal Stem Cells in a Type 1 Diabetes Mouse Model

“…Although an optimal treatment for acute or chronic graft-versus-host disease (GvHD) has not yet been developed, hMSCs have significant therapeutic potential. Since hMSCs promote immunosuppressive and immunomodulatory environments through a number of mechanisms, including protein/peptide/hormone secretion, mitochondrial delivery, and the transfer of exosomes or micelles containing RNA and other molecules, hMSCs are also considered a promising medium for cell therapy against T1D [22][23][24][25][26]. Although pancreatic insulin-producing β-like cells have been generated from MSCs in vitro and transplanted in diabetic animal models, their underlying mechanisms of action have not yet been elucidated [23].…”

“…Although pancreatic insulin-producing β-like cells have been generated from MSCs in vitro and transplanted in diabetic animal models, their underlying mechanisms of action have not yet been elucidated [23]. MSCs express chemokine receptors and play an important role in the homing effect [27] as well as in pancreatic regeneration [23,24]. Furthermore, the systemic administration of MSCs in STZ-induced diabetic rats resulted in an increase in β-cell volume [24,28].…”

“…MSCs express chemokine receptors and play an important role in the homing effect [27] as well as in pancreatic regeneration [23,24]. Furthermore, the systemic administration of MSCs in STZ-induced diabetic rats resulted in an increase in β-cell volume [24,28].…”