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Cardiac fibroblast paracrine factors modulate mouse embryonic stem cells 

SHEN Dan-Ya^1,2,3, CHEN Zong-Hai^1,4, ZHAO Ya-Nan^1,3, ZHOU Jian-Xia^1,3, GUAN Xiu-Wen⁵, LIANG Hua-Min^1,2,3,*

¹Department of Physiology, School of Basic Medicine, Huazhong University of Science and Technology, Wuhan 430030, China；²National Demonstration Center for Experimental Basic Medical Education, Huazhong University of Science and Technology, Wuhan 430030, China；³Hubei Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation; Huazhong University of Science and Technology, Wuhan 430030, China；⁴Department of Pathophysiology, Hubei University of Nationalities, Enshi 445000, China；⁵Department of General Surgery, Renmin Hospital of Huangpi District, Wuhan 430300, China

Abstract

The study aims to investigate the effects of cardiac fibroblast (CF) paracrine factors on murine embryonic stem cells (ESCs). Conditioned mediums from either neonatal cardiac fibroblasts (ConM-NCF) or adult cardiac fibroblasts (ConM-ACF) were diluted by 1:50 and 1:5, respectively, to investigate whether these conditioned mediums impact murine ESCs distinctly with RT-real time PCR techniques, cell proliferation essay, ELISA and by counting percentage of beating embryoid bodies (EBs) during ESCs differentiation. The data showed that the paracrine ability of CFs changed dramatically during development, in which interleukin 6 (IL6) increased with maturation. ConM-NCF 1:50 and ConM-NCF 1:5 had opposite effects on the pluripotent markers, although they both reduced mouse ESC proliferation. ConM-ACF 1:50 promoted ESCs pluripotent markers and proliferation, while ConM-ACF 1:5 exerted negative effects. All CF-derived conditioned mediums inhibited cardiac differentiation, but with distinguishable features: ConM-NCF 1:50 slightly decreased the early cardiac differentiation without altering the maturation tendency or cardiac specific markers in EBs at differentiation of day 17; ConM-ACF 1:50 had more significant inhibitory effects on early cardiac differentiation than ConM-NCF 1:50 and impeded cardiac maturation with upregulation of cardiac specific markers. In addition, IL6 neutralization antibody attenuated positive effect of ConM-ACF 1:50 on ESCs proliferation, but had no effects on ConM-NCF 1:50. Long-term IL6 neutralization reduced the percentage of beating EBs at early developmental stage, but did not alter the late cardiac differentiation. Taken together, both the quality and quantity of factors and cytokines secreted by CFs are critical for the ESC fate. IL6 could be a favorable cytokine for ESC pluripotency and the early cardiac differentiation.

Key words: murine embryonic stem cells; microenvironment; paracrine factors; cell therapy; interleukin 6

Received: 2020-03-16　　Accepted: 2020-08-17

Corresponding author: 梁华敏　　E-mail: hliang@mail.hust.edu.cn

DOI: 13294/j.aps.2020.0058

Citing This Article：

SHEN Dan-Ya, CHEN Zong-Hai, ZHAO Ya-Nan, ZHOU Jian-Xia, GUAN Xiu-Wen, LIANG Hua-Min. Cardiac fibroblast paracrine factors modulate mouse embryonic stem cells . Acta Physiol Sin 2020; 72 (5): 651-659