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Fig. 7 | BioMedical Engineering OnLine

Fig. 7

From: Wound healing of human embryonic stem cell-derived retinal pigment epithelial cells is affected by maturation stage

Fig. 7

Ca2+ wave propagation in wounded hESC-RPEs after mechanical stimulation. A bright field image of a hESC-RPE monolayer (9d + 7d) (A). The dark shadow of micropipette lowered towards the cell to perform mechanical stimulation can be seen. Fluorescence image of the same culture as in A loaded with fluorescent Ca2+ sensitive dye Fluo-4 AM that reflects [Ca2+]i concentration in the cytoplasm (B). A white line indicates the area of the monolayer that responded to single-cell mechanical stimulation. White dots indicate the cells that participate in a mechanically induced Ca2+ wave. Similarly a bright field image of a hESC-RPE monolayer (28 days + 7 days) (C). The dark shadow of micropipette lowered towards the cell to perform mechanical stimulation can be seen. Fluorescence image of the same culture as in A loaded with fluorescent Ca2+ sensitive dye Fluo-4 AM that reflects [Ca2+]i concentration in the cytoplasm (D). A white line indicates the area of the monolayer that responded to single-cell mechanical stimulation. White dots indicate the cells that participate in a mechanically induced Ca2+ wave. Number of hESC-RPE cells participating in a mechanically induced intercellular Ca2+ wave in wounded 9-day-cultured cells followed for 15 min (ctrl, ns = 7; wounded, ns = 7), or for 7 days (ctrl, ns = 7; wounded, ns = 7). The mechanically induced intercellular Ca2+ wave in wounded 28-day-cultured cells followed for 15 min (ctrl, ns = 8; wounded, ns = 8), or for 7 days (ctrl, ns = 6; wounded, ns = 7). E White bar—control; black bar—wound edge or healed wound. Statistical significance: p < 0.05 is indicated with Asterisk. Area of a mechanically induced intercellular Ca2+ wave spreading in wounded 9-day-cultured cells followed for 15 min (ctrl ns = 7; wounded, ns = 7), or for 7 days (ctrl, ns = 7; wounded, ns = 7). The mechanically induced intercellular Ca2+ wave spreading in wounded 28-day-cultured cells followed for 15 min (ctrl, ns = 8; wounded ns = 8), or for 7 days (ctrl, ns = 6; wounded, ns = 7). F White bar—control; black bar—wound edge or healed wound. Statistical significance: p < 0.05 is indicated with Asterisk. The presented data are combined from Regea08/017 and Regea08/023 hESC-RPEs

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