Role of TRPV1 in the Differentiation of Mouse Embryonic Stem Cells into Cardiomyocytes.

Yan Qi,Zhichao Li,Xiaoqiang Yao,Zenghua Qi,Yu Huang,Suk-Ying Tsang,Chun-Kit Wong,Iek-Chi Lo,Chun So,Ferenc Gallyas

doi:10.1371/journal.pone.0133211

Yan Qi, Zhichao Li + Show 8 more

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https://doi.org/10.1371/journal.pone.0133211

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Journal: PloS one	Publication Date: Jul 24, 2015
Citations: 22	License type: CC BY 4.0

Affiliation: Chinese University of Hong Kong

Abstract

Cytosolic Ca2+ ([Ca2+]i) is an important signal that regulates cardiomyocyte differentiation during cardiogenesis. TRPV1 is a Ca2+-permeable channel that is expressed in cardiomyocytes. In the present study, we utilized mouse embryonic stem cell-derived cardiomyocytes (mESC-CMs) as a model to investigate the functional role of TRPV1 in cardiomyocyte differentiation. Induction of embryonic stem cells into cardiomyocytes was achieved using embryoid body (EB)-based differentiation method. Quantitative PCRs showed an increased TRPV1 expression during the differentiation process. In [Ca2+]i measurement study, application of TRPV1 agonists, capsaicin and camphor, elicited a [Ca2+]i rise in mESC-CMs, the effect of which was abolished by TRPV1-shRNA. In functional study, treatment of EBs with TRPV1 antagonists (capsazepine and SB366791) and TRPV1-shRNA reduced the size of the EBs and decreased the percentage of spontaneously beating EBs. TRPV1 antagonists and TRPV1-shRNA also suppressed the expression of cardiomyocyte marker genes, including cardiac actin, c-TnT, c-TnI, and α-MHC. Taken together, this study demonstrated an important functional role of TRPV1 channels in the differentiation of mESCs into cardiomyocytes.

Highlights

Embryonic stem cells (ESCs) are derived from the inner cell mass of blastocysts
Immunostaining was used to examine the expression of TRPV1 proteins in mouse ESCs (mESCs) and mESC-CMs
The cells were stained on day 0, day 4 and day 9

Summary

Introduction

Embryonic stem cells (ESCs) are derived from the inner cell mass of blastocysts. They can selfrenew and are pluripotent, meaning that they can proliferate indefinitely and have the ability to differentiate into different cell lineages including cardiomyocytes [1]. ESC-derived cardiomyocytes (ESC-CMs) provide an unlimited ex vivo source of cardiomyocytes for cell-based heart therapies, and are an excellent model for studying heart development. Ultrastructural, molecular biological, and electrophysiological studies have demonstrated that in vitro differentiation of mESC-CMs within the EBs closely recapitulates the developmental.

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