CXCL4/PF4 is a predictive biomarker of cardiac differentiation potential of human induced pluripotent stem cells

Fumiya Ohashi,Yoshiki Sawa,Emiko Ito,Satoshi Yasuda,Takuya Kuroda,Yoji Sato,Masayoshi Itoh,Hideya Kawaji,Shigeru Miyagawa,Atsuhiro Saito,Shohei Yoshida,Tadashi Sameshima,Jun Kawai,Takumi Miura

doi:10.1038/s41598-019-40915-w

Abstract

Selection of human induced pluripotent stem cell (hiPSC) lines with high cardiac differentiation potential is important for regenerative therapy and drug screening. We aimed to identify biomarkers for predicting cardiac differentiation potential of hiPSC lines by comparing the gene expression profiles of six undifferentiated hiPSC lines with different cardiac differentiation capabilities. We used three platforms of gene expression analysis, namely, cap analysis of gene expression (CAGE), mRNA array, and microRNA array to efficiently screen biomarkers related to cardiac differentiation of hiPSCs. Statistical analysis revealed candidate biomarker genes with significant correlation between the gene expression levels in the undifferentiated hiPSCs and their cardiac differentiation potential. Of the candidate genes, PF4 was validated as a biomarker expressed in undifferentiated hiPSCs with high potential for cardiac differentiation in 13 additional hiPSC lines. Our observations suggest that PF4 may be a useful biomarker for selecting hiPSC lines appropriate for the generation of cardiomyocytes.

Highlights

Human induced pluripotent stem cells are capable of differentiating into various tissues[1], thereby acting as a source of cells for regenerative medicine and drug discovery[2,3,4,5,6,7,8]
We analysed the cardiomyocytes derived from human induced pluripotent stem cell (hiPSC) using flow cytometry, quantitative reverse transcription-polymerase chain reaction, immunostaining, and beating analysis, and determined the cardiac differentiation efficiency ranking
Our results suggest that the cardiac differentiation potential of individual hiPSC lines can be predicted by assessing PF4 expression in hiPSCs

Summary

Introduction

Human induced pluripotent stem cells (hiPSCs) are capable of differentiating into various tissues[1], thereby acting as a source of cells for regenerative medicine and drug discovery[2,3,4,5,6,7,8]. Several studies have suggested that the direction of differentiation of tissues derived from the endoderm, mesoderm, and ectoderm varies depending on the line of human embryonic stem cells (hESCs) and hiPSCs11–13. Variation in the direction of differentiation among hiPSC lines is the result of differences in somatic tissue of origin and epigenetic changes[14,15,16]. Biomarkers are required for selecting suitable hiPSC lines with high differentiation potential for specific tissues. We comprehensively analysed the gene expression of hiPSCs using cap analysis of gene expression (CAGE), mRNA array, and microRNA array to screen for biomarkers of cardiac differentiation potential. Our proposed method of using three gene analysis platforms for identifying novel predictive biomarkers of hiPSCs with high cardiac differentiation potential will identify useful genes that can be important for selecting desired hiPSC lines

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