Three-dimensional nanofiberous PLLA/PCL scaffold improved biochemical and molecular markers hiPS cell-derived insulin-producing islet-like cells

Nanofibrous scaffolds are considered as a new strategy for Type 1 diabetes mellitus therapy. We used a hybrid of poly-l-lactic acid (PLLA) and polycaprolactone (PCL) as three-dimensional (3D) culture models for differentiation of human induced pluripotent stem cells (hiPSCs) to beta islet-like cluster cell compared with routine culture (2D). Morphological changes of cells were checked by microscope. mRNA endodermal SOX-17 on day 7 and pancreatic gene markers Pdx1, glucagon and Glut2 were evaluated on day 23 by qPCR. As well as, insulin and C-peptide protein expression was evaluated by immunocytochemistry staining. In addition, insulin and C-peptide secretion in various glucose concentrations was evaluated by ELISA. Light and scanning electron microscopy (SEM) microscope showed changes in induced cells. In tandem, these modifications were more evident in 3 D culture. Pdx1, Glucagon and Glut2 markers in PLLA/PCL were significantly higher in 3 D culture. In addition, qualitative immunochemistry showed that insulin and C-peptide were expressed in 2 D and 3 D culture medium. Furthermore, evaluation of insulin and C-peptide clarified that secretion of these proteins in PLLA/PCL scaffold were statistically different in 2 D and 3 D strategies. These findings suggest that functional matured induction cells on PLLA/PCL scaffold can be used for islet beta cell therapy and regenerative medicine.