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Biomed Eng Online. 2018 Jul 31;17(1):102. doi: 10.1186/s12938-018-0535-z.

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

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Faculty of Biomedical Sciences and Engineering, BioMediTech, Tampere University of Technology, Arvo Ylpön Katu 34, Tampere, Finland.
Faculty of Medical and Life Sciences, BioMediTech, University of Tampere, Arvo Ylpön Katu 34, Tampere, Finland.
Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, Kuopio, Finland.
Department of Ophthalmology, Kuopio University Hospital, Kuopio, Finland.
Faculty of Medical and Life Sciences, BioMediTech, University of Tampere, Arvo Ylpön Katu 34, Tampere, Finland.



Wound healing of retinal pigment epithelium (RPE) is a complex process that may take place in common age-related macular degeneration eye disease. The purpose of this study was to evaluate whether wounding and wound healing has an effect on Ca2+ dynamics in human embryonic stem cell (hESC)-RPEs cultured different periods of time.


The 9-day-cultured or 28-day-cultured hESC-RPEs from two different cell lines were wounded and the dynamics of spontaneous and mechanically induced intracellular Ca2+ activity was measured with live-cell Ca2+ imaging either immediately or 7 days after wounding. The healing time and speed were analyzed with time-lapse bright field microscopy. The Ca2+ activity and healing speed were analysed with image analysis. In addition the extracellular matrix deposition was assessed with confocal microscopy.


The Ca2+ dynamics in hESC-RPE monolayers differed depending on the culture time: 9-day-cultured cells had higher number of cells with spontaneous Ca2+ activity close to freshly wounded edge compared to control areas, whereas in 28-day-cultured cells there was no difference in wounded and control areas. The 28-day-cultured, wounded and 7-day-healed hESC-RPEs produced wide-spreading intercellular Ca2+ waves upon mechanical stimulation, while in controls propagation was restricted. Most importantly, both wave spreading and spontaneous Ca2+ activity of cells within the healed area, as well as the cell morphology of 28-day-cultured, wounded and thereafter 7-day-healed areas resembled the 9-day-cultured hESC-RPEs.


This acquired knowledge about Ca2+ dynamics of wounded hESC-RPE monolayers is important for understanding the dynamics of RPE wound healing, and could offer a reliable functionality test for RPE cells. The data presented in here suggests that assessment of Ca2+ dynamics analysed with image analysis could be used as a reliable non-invasive functionality test for RPE cells.


Ca2+ waves; Cell maturation; Image analysis; Mechanical stimulation; Mechanically induced intercellular Ca2+ waves; RPE; Spontaneous [Ca2+]i increases; Wound healing; hESC-RPE

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