Adv Healthc Mater. 2019 May 14:e1900165. doi: 10.1002/adhm.201900165. [Epub ahead of print]

Clonal Isolation of Human Pluripotent Stem Cells on Nanofibrous Substrates Reveals an Advanced Subclone for Cardiomyocyte Differentiation.

Yu L^1,2, Li J^1,3, Minami I⁴, Qu X³, Miyagawa S³, Fujimoto N³, Hasegawa K¹, Chen Y^1,5, Sawa Y³, Kotera H^1,2, Liu L^1,3.

Abstract

Human pluripotent stem cells (hPSCs) have been widely used for various applications including disease modeling and regenerative medicine, among others. Recently, an increasing number of studies has focused on heterogeneity among hPSCs, which could affect cell quality and subsequent applications. In this study, a nanofibrous platform is developed for single human induced pluripotent stem cell isolation and culture. One type of single cell-derived subclone is established and found to have a distinct morphology compared to other subclones. When used for differentiation toward cardiomyocytes, this type of subclone demonstrates higher differentiation efficiency, increased maturation, and stronger beating compared to those derived from the other subclones. The findings provide a convenient method for single-cell isolation and culture, and demonstrate that variations in differentiation tendencies exist among subclones from the same cell line. This substrate adhesion-based selection process could be used to obtain cell lines with improved differentiation efficiency toward cardiomyocytes and other cell types, which would be advantageous for studies in various fields.