Fig. 1: HVPs expand, repopulate and functionally mature in an ex vivo 3D NHP heart model.
a, Schematic of the experimental setup for in vitro differentiation of HVPs from NKX2-5eGFP/wt hESCs (left) and their ex vivo co-culture with native NHP-LV slices in biomimetic chambers (right). b, Contractile force of ex vivo cultured NHP heart slices with and without HVPs on indicated days of co-culture. Box plot shows all data points as well as minimum, maximum, median and quartiles; n = 11 biological replicates per group; ***P < 0.001 (two-way ANOVA). c, Percentage of EdU+/eGFP+ and ClCasp3+/eGFP− cells during co-culture. Data are mean ± s.e.m.; n = 3 biological replicates per timepoint for EdU analysis; n = 4 biological replicates per timepoint for ClCasp3 analysis; *P < 0.05, **P < 0.005 versus D0 (one-way ANOVA). d,e, Left: representative immunofluorescence images of D50 chimeric human–NHP heart constructs using an antibody against GFP (a-GFP) together with antibodies for MLC2a and MLC2v (d) or CD31 (e). Scale bars, 100 µm (d), 50 µm (e) and 10 µm (insets). Right: percentage of eGFP+ cells expressing MLC2v, MLC2a or both (d) and human cells expressing CD31 (e) on D21 and D50. HuNu, human nuclear antigen. Data are mean ± s.e.m. and individual data points; n = 5 biological replicates per timepoint in d, n = 3 biological replicates per timepoint in e; *P < 0.05, **P < 0.005, ***P < 0.001 (two-way ANOVA for d and t-test for e). For b–e, exact P values and numerical data are provided in Source Data Fig. 1.
