In situ differentiation of human-induced pluripotent stem cells into functional cardiomyocytes on a coaxial PCL-gelatin nanofibrous scaffold

Sridharan, D and Palaniappan, A and Blackstone, BN and Dougherty, JA and Kumar, N and Seshagiri, PB and Sayed, N and Powell, HM and Khan, M (2021) In situ differentiation of human-induced pluripotent stem cells into functional cardiomyocytes on a coaxial PCL-gelatin nanofibrous scaffold. In: Materials Science and Engineering C, 118 .

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Abstract

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

Item Type:	Journal Article
Publication:	Materials Science and Engineering C
Publisher:	Elsevier Ltd
Additional Information:	The copyright of this article belongs to Elsevier Ltd
Keywords:	Efficiency; Fluorescence; Fourier transform infrared spectroscopy; Gene expression; Heart; Nanofibers; Scanning electron microscopy; Stem cells, Cardiac differentiation; Cardiac progenitors; Cardiac regeneration; Gene expression analysis; Gene Expression Data; Human-induced pluripotent stem cells; Immunofluorescence staining; Nanofibrous scaffolds, Scaffolds (biology)
Department/Centre:	Division of Biological Sciences > Molecular Reproduction, Development & Genetics
Date Deposited:	07 Oct 2020 11:25
Last Modified:	07 Oct 2020 11:25
URI:	http://eprints.iisc.ac.in/id/eprint/66589

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