Wound healing of human embryonic stem cell-derived retinal pigment epithelial cells is affected by maturation stage
| Autor: | Kai Kaarniranta, Amna E. Abu Khamidakh, Kati Juuti-Uusitalo, Alejandra Rodriguez-Martinez, Jari Hyttinen, Anne Kallioniemi, Heli Skottman |
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| Přispěvatelé: | Lääketieteen ja biotieteiden tiedekunta - Faculty of Medicine and Life Sciences, University of Tampere, Tampere University, Faculty of Biomedical Sciences and Engineering, Faculty of Medicine and Life Sciences, Research group: Computational Biophysics and Imaging Group |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Mechanically induced intercellular Ca2+ waves lcsh:Medical technology Human Embryonic Stem Cells Biomedical Engineering Wound healing Retinal Pigment Epithelium Cell morphology Image analysis Biokemia solu- ja molekyylibiologia - Biochemistry cell and molecular biology Biomaterials Extracellular matrix 03 medical and health sciences chemistry.chemical_compound hESC-RPE medicine Humans Radiology Nuclear Medicine and imaging 3125 Otorhinolaryngology ophthalmology Spontaneous [Ca2+]i increases Mechanical stimulation Retinal pigment epithelium Radiological and Ultrasound Technology Ca2+ waves Chemistry Research Cell Differentiation Retinal 217 Medical engineering General Medicine Embryonic stem cell Mechanical stimulation Ca2+ waves Cell biology 030104 developmental biology medicine.anatomical_structure lcsh:R855-855.5 Cell culture Lääketieteen tekniikka - Medical engineering 1182 Biochemistry cell and molecular biology RPE Cell maturation Intracellular |
| Zdroj: | BioMedical Engineering OnLine, Vol 17, Iss 1, Pp 1-20 (2018) BioMedical Engineering |
| Popis: | Background 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. Methods 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. Results 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. Conclusions 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. Electronic supplementary material The online version of this article (10.1186/s12938-018-0535-z) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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