Dataset Information

Effect of migratory behaviors on human induced pluripotent stem cell colony formation on different extracellular matrix proteins.

ABSTRACT:

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.

SUBMITTER: Chang J

PROVIDER: S-EPMC6260426 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Publications

Effect of migratory behaviors on human induced pluripotent stem cell colony formation on different extracellular matrix proteins.

Chang Jessica J Kim Mee-Hae MH Agung Eviryanti E Senda Sho S Kino-Oka Masahiro M

Regenerative therapy 20181120

<h4>Introduction</h4>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.<h4>Methods</h4>To quantify how different ECM proteins affect migratory behavior during the colony formation process, single cells were seeded ...[more]

PMID: 30525068

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Project description:Diffuse infiltration across brain tissue is a hallmark of glioblastoma and the main cause of unsuccessful total resection that leads to tumor reappearance. A subpopulation termed glioblastoma stem cells (GSCs) has been directly related to aggressive invasion; nonetheless, their migratory characteristics and regulation by the microenvironment are still unknown. In this study, we developed a composite matrix of hyaluronan (HA) structurally supported by a collagen-oligomer fibril network to simulate the brain tumor extracellular matrix (ECM) composition. Matrigel-coated microfibers were embedded within the matrix to create a tunable dual niche microenvironment that resembles the vascular network of the brain. This model was compared with the most commonly used in vitro three-dimensional (3D) culture formats, Matrigel and collagen type-I monomer matrices, to study how the mechanical and compositional properties of the ECM alter the migration characteristics of GSC neurospheres. The migration mode, distance, velocity, and morphology of the GSCs were monitored over a 72-h period. The cells altered their migration mode depending on the matrix composition, showing migration by expansive growth in Matrigel matrices, multicellular extension along rigid interfaces (as Matrigel glass and coated microfibers), and mesenchymal single-cell migration in collagen matrices. Velocity and distance of migration within each composition varied according to matrix mechanical properties. In the dual niche system, the presence of HA reduced velocity and number of migratory cells; however, cells that came in contact with the pseudovessels exhibited collective migration by an extensive strand and reached higher velocities than cells migrating individually across the 3D matrix. Our results show that GSCs adopt varied migration mechanisms to invade multiple ECM microenvironments, and the migration characteristics exhibited are highly influenced by the matrix physical properties. Moreover, GSC neurospheres exhibit concomitant single and collective migration as a function of the microenvironment topography to reach the most productive migration strategy.

Dataset Information

Effect of migratory behaviors on human induced pluripotent stem cell colony formation on different extracellular matrix proteins.

Introduction

Methods

Results

Conclusions

Publications

Effect of migratory behaviors on human induced pluripotent stem cell colony formation on different extracellular matrix proteins.

Similar Datasets

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