Extended Data Fig. 3: Generation and analysis of an acute ex-vivo NHP heart injury model.
a, Standardized non-transmural myocardial injury in NHP heart slices by defined RFA. Live and dead cells are stained by calcein and ethidium dimer, respectively (middle) Scale bar 200 µm. Intact ECM scaffold after RFA injury shown by WGA immunostaining. Scale bar 200 µm. Stainings were performed immediately after RFA. b, Representative fluorescence images of RFA-injured slices after immunostaining for Collagen type I (ColI) combined with cTNT (top) or DDR2 (bottom) on indicated days. Lower panels show images of the RFA area. Scale bars 30 µm (top) and 25 µm (bottom). c, Sequential live imaging of NKX2-5eGFP/wt HVPs migrating from the seeding frame into the tissue showing homogenous repopulation of the slice by D15 in the absence of RFA injury. Scale bars 200 µm. d, FACS analysis of HVP and CM batches before seeding indicating cellular purity (ISL-1, cTNT and TRA-1-60). Data are mean ± SEM and individual data points, n = 3 biological replicates per group, ***p < 0.001 vs HVPs (two-way ANOVA). e, Representative immunostainings of eGFP and cTNT in RFA-injured area on D14 after selective seeding of NKX2-5eGFP/wt HVPs (left) or CMs (right). Scale bars 50 µm. f, Two-photon live microscopy of RFA-injured slices for eGFP and second-harmonic-imaging (SHG) visualization of collagen and scar size on D21. Circles demarcate areas with collagen deposition. Scale bars 100 µm. g, Trans-well migration assays with D0 NKX2-5eGFP/wt HVPs in the upper and NHP heart slice in the lower compartment, respectively. Images show trans-well migrated HVPs on polycarbonate membrane in the absence (left) or presence of multiple (middle) or single (right) RFA injury. Dashed line marks the site of HVP accumulation. Scale bars 2 mm. h, Trans-well migration assays with D0 NKX2-5eGFP/wt HVPs in the upper and decellularized NHP heart slice in the lower compartment with a single RFA injury. Scale bar 2 mm. For d, exact p-values and numerical data are provided in Source Data Extended Data Fig. 3.
