Semi-automatic Method for Ca2+ Imaging Data Analysis of Maturing Human Embryonic Stem Cells-Derived Retinal Pigment Epithelium

Amna E. Abu Khamidakh, Florentino Caetano Dos Santos, Heli Skottman, Kati Juuti-Uusitalo, Jari Hyttinen

    Research output: Contribution to journalArticleScientificpeer-review

    4 Citations (Scopus)

    Abstract

    Ca2+ is a second messenger controlling vital cellular processes, including cell maturation. Changes in Ca2+ signaling during maturation of human embryonic stem cell-derived retinal pigment epithelial cells (hESC-RPE) have not been assessed previously. The aim of this study was to investigate maturation-dependent changes in transient intracellular Ca2+ ([Ca2+]i) increases in hESC-RPE. For this, we developed image analysis tools to evaluate cell-specific Ca2+ signals from the entire field of view. Spontaneous and mechanically induced transient [Ca2+]i increases (STIs and MITIs) were analyzed in hESC-RPEs cultured for 9 or 28 days, altogether from more than 80,000 cells. Both cultures showed STIs: the longer culture time resulted in twofold increase of amount of cells with STIs. Mechanical stimulation induced intercellular Ca2+ waves in cells from both time points, but longer culture time reduced Ca2+ wave spreading. Depletion of intracellular Ca2+ stores decreased cell fraction with STIs and MITIs at both time points, and absence of extracellular Ca2+ had similar effect on cells with STIs. To conclude, hESC-RPE cells undergo significant Ca2+ signaling re-arrangements during a short maturation period increasing cell fraction with STIs, while decreasing coordinated cell response to mechanical stimulation. This knowledge and proposed analysis tools can be used for assessment of hESC-RPE maturation in vitro.

    Original languageEnglish
    Pages (from-to)1-13
    Number of pages13
    JournalAnnals of Biomedical Engineering
    Volume44
    Issue number11
    DOIs
    Publication statusPublished - 2016
    Publication typeA1 Journal article-refereed

    Keywords

    • Calsium wave
    • Intercellular calcium waves
    • Mechanical stimulation
    • Mechanically induced transient [Ca] increase
    • Retinal pigment epithelial cell
    • Spontaneous transient [Ca] increase
    • cell maturation
    • hESC-RPE
    • image analysis

    Publication forum classification

    • Publication forum level 2

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