Figure 3.

Differentiation potential of ciCPCs under in vitro conditions

(A) Schematic of cell expansion and differentiation procedures. (B) Heatmap of cardiovascular genes detected by qPCR in TTFs, early ciCPCs, late ciCPCs, and differentiated cells (DCs). (C) Immunostaining of sarcomeric α-Actinin and Cx43 in ciCPCs treated by CM differentiation medium. Scale bars, 50 μm. (D) Representative Ca²⁺ waves detected by Fluo-4 dye in ciCPC-CMs at Ca²⁺ maximum and minimum using live-cell imaging. Fluorescent images correspond to the Video S3. Scale bar, 20 μm. (E) Representative traces of action potentials in ciCPC-CMs (18 cell beating clusters) analyzed by the intracellular electrical recording. (F) Immunostaining of αSMA and SM-MHC in ciCPCs treated by differentiation medium. Scale bars, 100 μm. (G) Corresponding I-V plot of ciCPC-SMCs in response to the vehicle (n = 5), nifedipine (n = 5), or FPL641176 (n = 7) as detected by the whole-cell patch-clamp. Current is given in pA/pF and membrane potential in mV. Data measures are presented as mean ± standard error. (H) Immunostaining of CD31 and vWF in ciCPCs treated by differentiation medium. Scale bars, 100 μm. (I) In vitro tube formation potential of ciCPC-ECs analyzed on Matrigel. Scale bar, 500 μm. See also Figure S4.