Tóm tắt

Human-induced pluripotent stem cells (hiPSCs) derived cardiomyocytes (hiPSC-CMs) have been explored for rncardiac regeneration and repair as well as for the development of in vitro 3D cardiac tissue models. Existing rnprotocols for cardiac differentiation of hiPSCs utilize a 2D culture system. However, the efficiency of hiPSC rndifferentiation to cardiomyocytes in 3D culture systems has not been extensively explored. In the present study, rnwe investigated the efficiency of cardiac differentiation of hiPSCs to functional cardiomyocytes on 3D nanofi-brous scaffolds. Coaxial polycaprolactone (PCL)-gelatin fibrous scaffolds were fabricated by electrospinning and rncharacterized using scanning electron microscopy (SEM) and fourier transform infrared (FTIR) spectroscopy. rnhiPSCs were cultured and differentiated into functional cardiomyocytes on the nanofibrous scaffold and com-pared with 2D cultures. To assess the relative efficiencies of both the systems, SEM, immunofluorescence staining rnand gene expression analyses were performed. Contractions of differentiated cardiomyocytes were observed in rn2D cultures after 2 weeks and in 3D cultures after 4 weeks. SEM analysis showed no significant differences in the rnmorphology of cells differentiated on 2D versus 3D cultures. However, gene expression data showed significantly rnincreased expression of cardiac progenitor genes (ISL-1, SIRPA) in 3D cultures and cardiomyocytes markers rn(TNNT, MHC6) in 2D cultures. In contrast, immunofluorescence staining showed no substantial differences in rnthe expression of NKX-2.5 and α-sarcomeric actinin. Furthermore, uniform migration and distribution of the in rnsitu differentiated cardiomyocytes was observed in the 3D fibrous scaffold. Overall, our study demonstrates that rncoaxial PCL-gelatin nanofibrous scaffolds can be used as a 3D culture platform for efficient differentiation of rnhiPSCs to functional cardiomyocytes.