The Different Biological Characteristics Of Embryonic Epicardial Progenitor Cells In Vitro

Part one Isolation and culture of different embryonic day epicardial progenitor cellsThe embryonic days of excised embryonic mouse ventricles used to isolate epicardial progenitor cells(EpiCs) are not uniform, and embryonic day(E)10.5-E13.5 embryonic ventricles are used to isolate EpiCs.Objective: To explore how to successfully culture different embryonic day epicardial progenitors cells.Methods: We chose E11.5, E13.5, and E15.5 embryonic ventricle explants to isolate EpiCs. We used PCR and immunofluorescence to confirm their epicardial identity.Results: We successfully cultured EpiCs from E11.5, E13.5, and E15.5 embryonic ventricle explants. E11.5, E13.5, and E15.5 EpiC isolation out of the explants required different time windows.Conclusions: These results suggest that different embryonic stages of ventricle should be properly chosen for culturing EpiCs according to the purposes of various experiments.Part two Experimental study of the differences of different embryonic day EpiC biological characteristicsFrom this early formation of the epicardium until adulthood,epicardial cells undergo dynamic changes in the expression of embryonic genes that correlate with changes in the epicardial cell properties. Previous studies have not examined the differences in the biological characteristics of EpiCs on different embryonic days, and the differences maybe affect the related results of experiments that use EpiCs.Objective: To explore the differences of different embryonic day EpiC biological characteristics.Methods: The differences in the migration, proliferation and differentiation of different embryonic day EpiCs were studied by scratch assay, cell cycle analysis and platelet derived growth factor-bb(PDGF-BB)treatment.Results: The migratory abilities of E11.5, E13.5, and E15.5 EpiCs decreased during embryonic development; however, no difference in the proliferative abilities of these cells was generally observed. Early stage EpiCs were more prone to express smooth muscle cell markers than later stage EpiCs, so the progenitor cell characteristics of early stage EpiCs was better than those of later stage EpiCs.Conclusions: Different embryonic day EpiCs have different biological characteristics. To avoid the influence of the different biologicalcharacteristics of these cells on different embryonic days, the same experiment should choose the same stage of embryonic ventricle explants to culture EpiCs.