Fig. 1
Generation and characterization of BCP. a Representative images of the process of decellularization of rat placenta. (I) Native placenta before decellularization; (II) after perfusion with 1%SDS for 1 h; (III) after perfusion with 1%SDS for 18 h; (IV) after perfusion with 1%SDS for 24 h. b Hematoxylin and eosin (H&E), Sirius Red, and DAPI/cTnT staining of sections from the native rat placenta (left), DP (Middle), and BCP (Right). The staining indicated that the cellular component and visible nuclei were removed successfully from the placenta, and DP scaffold was recellularizated with iPSC-CMs to generate a BCP. Scanning electron micrographs showed the composite of materials and network in the DP scaffold with high porosity and recellularized with hiPSC-CMs after seeding in the BCP. c (A) The activation map of BCP was evaluated after 7 days by staining with a Ca2+-sensitive dye (5 μM fura-2, AM), then measurement of the intensity of transmitted light (interval: 8 ms, sampling time:2 ms/frame). Bar, 500 μm. (B) Representative optical traces of intracellular Ca2+ transient at the intrinsic rate and electrical pacing at a rate of 40 times/min, 60 times/min, 90 times/min, 100 times/min, and with Isoprenaline 10 μm/ml perfusion (x-axis: time, y-axis: dF/F). d Quantification of multiple rat pro-survival and/or stem cell-recruiting factors (Activin A, brain-derived neurotropic factor [BDNF], beta-nerve growth factor [bNGF], glial cell-derived neurotrophic factor [GDNF], Granulocyte-macrophage colony-stimulating factor [GM-CSF] and insulin-like growth factor binding protein 5 [IGFBP-5]); and pro-angiogenic factors (basic fibroblast growth factor [bFGF], hepatocyte growth factor [HGF], insulin-like growth factor 1 [IGF-1], platelet-derived growth factor [PDGF]-AA, PDGF-BB and vascular endothelial growth factor [VEGF]) in the native rat placenta, DP and BCP. Error bars represent mean ± SEM of four independent experiments. Comparisons of native placenta, DP and BCP were performed using one-way ANOVA followed by Tukey’s post hoc test. *P < 0.05 compare with native placenta