Fig. 2: Medium conditioned by placental tissue and differentiated trophoblasts promotes fetal heart development and cardiomyocyte growth.
From: Placental extracellular vesicles promote cardiomyocyte maturation and fetal heart development
a Schematic indicating four culture conditions: fetal hearts cultured in unconditioned media (CTRL), fetal hearts cultured in media conditioned by E15.5 placental tissue (Placental CM), fetal hearts cultured in media conditioned by 5-day differentiated Ovol2+/+ TS cells (Trophoblast CM), and fetal hearts cultured in media conditioned by 5-day differentiated Ovol2-/- TS cells (Ovol2-Null CM). b Average number of beats per minute when fetal hearts were exposed to CTRL, Placental CM, Trophoblast CM, or Ovol2-Null CM for 24 and 48 h. c Quantification of epicardial cell outgrowth from fetal hearts exposed to CTRL, Placental CM, Trophoblast CM, or Ovol2-Null CM for 24 and 48 h. c Representative images of epicardial cell outgrowths at 24 h. A red dashed line delineates fetal heart borders and arrows demarcate examples of outgrowths. d Expression of six genes (Tead1, Myh7, Myl7, Acta2, Nppa, and Nppb) associated with cardiac development in cardiomyocytes exposed to CTRL, Placental CM, Trophoblast CM, or Ovol2-Null CM. e Western blot showing levels of Troponin T (TnT) and NKX2-5 in cardiomyocytes treated with CTRL, Placental CM, Trophoblast CM, or Ovol2-Null CM. ACTB was used as a loading control. Quantification of relative band intensity is shown to the right of the representative blots. f Immunofluorescence for TnT (green) in cardiomyocytes treated with CTRL, Placental CM, Trophoblast CM, or Ovol2-Null CM. DAPI was used to demarcate nuclei. Values significantly different from CTRL (N = 6 biologically independent samples, P < 0.05) are denoted with an asterisk (*); values significantly increased from CTRL but decreased from Placental CM are denoted with a number sign (#). One-way ANOVA followed by Tukey’s post-hoc test was used to determine statistical significance. Scale bar = 100 µm. Graphs represent means ± SEM. Uncropped images of western blots are provided in Supplementary Fig. 8.
