Angiotensin II promotes differentiation of mouse c-kit-positive cardiac stem cells into pacemaker-like cells

Xue,Cheng; Zhang,Jun; Lv,Zhan; Liu,Hui; Huang,Congxin; Yang,Jing; Wang,Ten

doi:10.3892/mmr.2015.3149

Angiotensin II promotes differentiation of mouse c-kit-positive cardiac stem cells into pacemaker-like cells

Authors:
- Cheng Xue
- Jun Zhang
- Zhan Lv
- Hui Liu
- Congxin Huang
- Jing Yang
- Ten Wang
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Affiliations: Department of Cardiology, The Affiliated Hospital of North Sichuan Medical College, Nanchong, Sichuan 637000, P.R. China, Department of Cardiology, Renmin Hospital of Wuhan University, Cardiovascular Research Institute, Wuhan University, Wuhan, Hubei 430060, P.R. China
Published online on: January 7, 2015 https://doi.org/10.3892/mmr.2015.3149
Pages: 3249-3258
Copyright: © Xue et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Cardiac stem cells (CSCs) can differentiate into cardiac muscle‑like cells; however, it remains unknown whether CSCs may possess the ability to differentiate into pacemaker cells. The aim of the present study was to determine whether angiotensin II (Ang II) could promote the specialization of CSCs into pacemaker‑like cells. Mouse CSCs were treated with Ang II from day 3‑5, after cell sorting. The differentiation potential of the cells was then analyzed by morphological analysis, flow cytometry, reverse transcription‑polymerase chain reaction, immunohistochemistry and patch clamp analysis. Treatment with Ang II resulted in an increased number of cardiac muscle‑like cells (32.7±4.8% vs. 21.5±4.8%; P<0.05), and inhibition of smooth muscle‑like cells (6.2±7.3% vs. 20.5±5.1%; P<0.05). Following treatment with Ang II, increased levels of the cardiac progenitor‑specific markers GATA4 and Nkx2.5 were observed in the cells. Furthermore, the transcript levels of pacemaker function‑related genes, including hyperpolarization‑activated cyclic nucleotide‑gated (HCN)2, HCN4, T‑box (Tbx)2 and Tbx3, were significantly upregulated. Immunofluorescence analysis confirmed the increased number of pacemaker‑like cells. The pacemaker current (If) was recorded in the cells derived from CSCs, treated with Ang II. In conclusion, treatment of CSCs with Ang II during the differentiation process modified cardiac‑specific gene expression and resulted in the enhanced formation of pacemaker‑like cells.

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