Cardiac fibroblast paracrine factors modulate mouse embryonic stem cells / 生理学报

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 150 and 15, 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 150 and ConM-NCF 15 had opposite effects on the pluripotent markers, although they both reduced mouse ESC proliferation. ConM-ACF 150 promoted ESCs pluripotent markers and proliferation, while ConM-ACF 15 exerted negative effects. All CF-derived conditioned mediums inhibited cardiac differentiation, but with distinguishable features ConM-NCF 150 slightly decreased the early cardiac differentiation without altering the maturation tendency or cardiac specific markers in EBs at differentiation of day 17; ConM-ACF 150 had more significant inhibitory effects on early cardiac differentiation than ConM-NCF 150 and impeded cardiac maturation with upregulation of cardiac specific markers. In addition, IL6 neutralization antibody attenuated positive effect of ConM-ACF 150 on ESCs proliferation, but had no effects on ConM-NCF 150. 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.