Differentiation of Alginate-Encapsulated Wharton Jelly-Derived Mesenchymal Stem Cells into Insulin Producing Cells

Insulin insufficiency due to the reduced pancreatic beta cell number is the hallmark of diabetes, resulting inan intense focus on the development of beta-cell replacement options. One approach to overcome the problem is tosearch for alternative sources to induce insulin-producing cells (IPCs), the advent of mesenchymal stem cells (MSCs)holds great promise for producing ample IPCs. Encapsulate the MSCs with alginate improved anti-inflammatory effectsof MSC treatment. This study aimed to evaluate the differentiation of wharton jelly-derived MScs into insulin producingcells using alginate encapsulation. In this experimental study, we established an efficient IPCs differentiation strategy of humanMSCs derived from the umbilical cord’s Wharton jelly with lentiviral transduction of Pancreas/duodenum homeoboxprotein 1 (PDX1) in a 21-day period using alginate encapsulation by poly-L-lysine (PLL) and poly-L-ornithine (PLO)outer layer. During differentiation, the expression level of PDX1 and secretion of insulin proteins were increased. Results showed that during time, the cell viability remained high at 87% at day 7. After 21 days, the differentiated beta-like cells in microcapsules were morphologically similar to primary beta cells. Evaluation of the expression of PDX1 and INS by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) on days 7, 14 and 21 of differentiation exhibited the highest expression on day 14. At the protein level, the expression of these two pancreatic markers was observed after PDX1 transduction. Results showed that the intracellular and extracellular insulin levels in the cells receiving PDX1 is higher than the control group. The current data showed that encapsulation with alginate by PLL and PLO outer layer permitted to increase the microcapsules’ beta cell differentiation. Encapsulate the transduced-MSCs with alginate can be applied in an in vivo model in order to do the further analysis.