Marek KukumbergTatsanee PhermthaiSuparat WichitwiengratXiaoyuan WangSubramanian ArjunanSuet Yen ChongChui Yee FongJiong Wei WangAbdul Jalil RufaihahCitra Nurfarah Zaini MattarNational University Health SystemNUS Yong Loo Lin School of MedicineFaculty of Medicine Siriraj Hospital, Mahidol UniversityNational Heart Centre, Singapore2022-08-042022-08-042021-12-01Scientific Reports. Vol.11, No.1 (2021)204523222-s2.0-85098949058https://repository.li.mahidol.ac.th/handle/123456789/79281Secretome derived from human amniotic fluid stem cells (AFSC-S) is rich in soluble bioactive factors (SBF) and offers untapped therapeutic potential for regenerative medicine while avoiding putative cell-related complications. Characterization and optimal generation of AFSC-S remains challenging. We hypothesized that modulation of oxygen conditions during AFSC-S generation enriches SBF and confers enhanced regenerative and cardioprotective effects on cardiovascular cells. We collected secretome at 6-hourly intervals up to 30 h following incubation of AFSC in normoxic (21%O2, nAFSC-S) and hypoxic (1%O2, hAFSC-S) conditions. Proliferation of human adult cardiomyocytes (hCM) and umbilical cord endothelial cells (HUVEC) incubated with nAFSC-S or hAFSC-S were examined following culture in normoxia or hypoxia. Lower AFSC counts and richer protein content in AFSC-S were observed in hypoxia. Characterization of AFSC-S by multiplex immunoassay showed higher concentrations of pro-angiogenic and anti-inflammatory SBF. hCM demonstrated highest proliferation with 30h-hAFSC-S in hypoxic culture. The cardioprotective potential of concentrated 30h-hAFSC-S treatment was demonstrated in a myocardial ischemia–reperfusion injury mouse model by infarct size and cell apoptosis reduction and cell proliferation increase when compared to saline treatment controls. Thus, we project that hypoxic-generated AFSC-S, with higher pro-angiogenic and anti-inflammatory SBF, can be harnessed and refined for tailored regenerative applications in ischemic cardiovascular disease.Mahidol UniversityMultidisciplinaryArticleSCOPUS10.1038/s41598-020-80326-w