Effect of migratory behaviors on human induced pluripotent stem cell colony formation on different extracellular matrix proteins
| Autor: | Mee-Hae Kim, Jessica Chang, Sho Senda, Masahiro Kino-oka, Eviryanti Agung |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Biomedical Engineering Clonal colony formation Biomaterials Extracellular matrix Focal adhesion 03 medical and health sciences 0302 clinical medicine Cell–cell interaction Cytoskeletal formation Clonal colony Cell migration lcsh:QH573-671 Induced pluripotent stem cell lcsh:R5-920 lcsh:Cytology Chemistry Human induced pluripotent stem cells Actin cytoskeleton Cell biology 030104 developmental biology Cell culture Original Article lcsh:Medicine (General) 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Regenerative Therapy Regenerative Therapy, Vol 10, Iss, Pp 27-35 (2019) |
| ISSN: | 2352-3204 |
| DOI: | 10.1016/j.reth.2018.10.004 |
| Popis: | Introduction Understanding how extracellular matrix (ECM) protein composition regulates the process of human induced pluripotent stem cell (hiPSC) colony formation may facilitate the design of optimal cell culture environments. In this study, we investigated the effect of migratory behaviors on hiPSC colony formation on various ECM-coated surfaces. Methods To quantify how different ECM proteins affect migratory behavior during the colony formation process, single cells were seeded onto surfaces coated with varying concentrations of different ECM proteins. Cell behavior was monitored by time-lapse observation, and quantitative analysis of migration rates in relation to colony formation patterns was performed. Actin cytoskeleton, focal adhesions, and cell–cell interactions were detected by fluorescence microscopy. Results Time-lapse observations revealed that different mechanisms of colony formation were dependent upon the migratory behavior of cells on different ECM surfaces. HiPSCs formed tight colonies on concentrated ECM substrates, while coating with dilute concentrations of ECM yielded more motile cells and colonies capable of splitting into single cells or small clusters. Enhanced migration caused a reduction of cell–cell contacts that enabled splitting or merging between cells and cell clusters, consequently reducing the efficiency of clonal colony formation. High cell-to-cell variability in migration responses to ECM surfaces elicited differential focal adhesion formation and E-cadherin expression within cells and colonies. This resulted in variability within focal adhesions and further loss of E-cadherin expression by hiPSCs. Conclusions Migration is an important factor affecting hiPSC colony-forming patterns. Regulation of migratory behavior can be an effective way to improve the expansion of hiPSCs while improving the process of clonal colony formation. We believe that this investigation provides a valuable method for understanding cell phenotypes and heterogeneity during colony formation in culture. Highlights • hiPSC colony-forming patterns were dependent on migratory behavior on different ECM surfaces. • Colony formation without splitting during migration improved efficiency of clonal colony formation. • Variability in migration behavior elicited differential cytoskeletal formation and E-cadherin expression. • Our method is valuable for understanding cell phenotypes and heterogeneity during colony formation. |
| Databáze: | OpenAIRE |
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