The human amniotic fluid stem cell secretome triggers intracellular Ca oscillations, NF-κB nuclear translocation and tube formation in human endothelial colony-forming cells.

Second trimester foetal human amniotic fluid-derived stem cells (hAFS) have been shown to possess remarkable cardioprotective paracrine potential in different preclinical models of myocardial injury and drug-induced cardiotoxicity. The hAFS secretome, namely the total soluble factors released by cells in their conditioned medium (hAFS-CM), can also strongly sustain in vivo angiogenesis in a murine model of acute myocardial infarction (MI) and stimulates human endothelial colony-forming cells (ECFCs), the only truly recognized endothelial progenitor, to form capillary-like structures in vitro. Preliminary work demonstrated that the hypoxic hAFS secretome (hAFS-CM ) triggers intracellular Ca oscillations in human ECFCs, but the underlying mechanisms and the downstream Ca -dependent effectors remain elusive. Herein, we found that the secretome obtained by hAFS undergoing hypoxic preconditioning induced intracellular Ca oscillations by promoting extracellular Ca entry through Transient Receptor Potential Vanilloid 4 (TRPV4). TRPV4-mediated Ca entry, in turn, promoted the concerted interplay between inositol-1,4,5-trisphosphate- and nicotinic acid adenine dinucleotide phosphate-induced endogenous Ca release and store-operated Ca entry (SOCE). hAFS-CM -induced intracellular Ca oscillations resulted in the nuclear translocation of the Ca -sensitive transcription factor p65 NF-κB. Finally, inhibition of either intracellular Ca oscillations or NF-κB activity prevented hAFS-CM -induced ECFC tube formation. These data shed novel light on the molecular mechanisms whereby hAFS-CM induces angiogenesis, thus providing useful insights for future therapeutic strategies against ischaemic-related myocardial injury.

© 2021 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.