A human induced pluripotent stem (hiPS) cell model for the holistic study of epithelial to mesenchymal transitions (EMTs)

Hookway, C.; Borensztejn, A.; Harris, L. K.; Carlson, S.; Dalgin, G.; Mishra, S.; Nivedita, N.; Adams, E. M.; Barszczewski, T.; Dixon, J. C.; Edmonds, J. H.; Ehlers, E. A.; Ferrante, A. J.; Fuqua, M. A.; Garrison, P.; Gopalan, J.; Gregor, B. W.; Hedayati, M. J.; Klein, K. N.; Leveille, C. L.; Meharry, S. L.; Morris, H. S.; Nadarajan, G.; Oluoch, S. A.; Parent, S. E.; Phan, A.; Roberts, B.; Sanchez, E. E.; Sluzewski, M. F.; Snyder, L. S.; Thirstrup, D. J.; Thottam, J. P.; Torvi, J. R.; Turman, G.; Viana, M. P.; Wilhelm, L.; Wijesooriya, C. S.; Yao, J.; Theriot, J. A.; Rafelski, S. M.; Gunawarda

doi:10.1101/2024.08.16.608353

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bioRxiv · 10.1101/2024.08.16.608353

A human induced pluripotent stem (hiPS) cell model for the holistic study of epithelial to mesenchymal transitions (EMTs)

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Abstract

The epithelial to mesenchymal transition (EMT) is a widely studied but poorly defined state change due to the variety of ways in which it has been characterized in cells. There is a need for reproducible cell model systems that enable the integration and comparison of different types of measured observations of cells across many distinct cellular contexts. We present human induced pluripotent stem (hiPS) cells as such a model system by demonstrating its utility through a comparative analysis of hiPS cell-EMT in 2D and 3D cell culture geometries. We developed live-imaging-based assays to directly compare examples of changes in cell function (via migration timing), molecular components (via expression of marker proteins), organization (via reorganization of cell junctions), and environment (via dynamics of basement membrane) in the same experimental system. The EMT-related changes we measured occurred earlier in 2D colonies than in 3D lumenoids, likely due to differences in the basement membrane environments associated with 2D vs. 3D initial hiPS cell culture geometries. We have made the 449 60-hour-long 3D time-lapse movies and the associated tools used for analysis and visualization open-source and easily accessible as a resource for future work in this field.

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Hookway, C., Borensztejn, A., Harris, L. K., Carlson, S., Dalgin, G., Mishra, S., Nivedita, N., Adams, E. M., Barszczewski, T., Dixon, J. C., Edmonds, J. H., Ehlers, E. A., Ferrante, A. J., Fuqua, M. A., Garrison, P., Gopalan, J., Gregor, B. W., Hedayati, M. J., Klein, K. N., Leveille, C. L., Meharry, S. L., Morris, H. S., Nadarajan, G., Oluoch, S. A., Parent, S. E., Phan, A., Roberts, B., Sanchez, E. E., Sluzewski, M. F., Snyder, L. S., Thirstrup, D. J., Thottam, J. P., Torvi, J. R., Turman, G., Viana, M. P., Wilhelm, L., Wijesooriya, C. S., Yao, J., Theriot, J. A., Rafelski, S. M., Gunawarda. 2024-08-19. A human induced pluripotent stem (hiPS) cell model for the holistic study of epithelial to mesenchymal transitions (EMTs). https://doi.org/10.1101/2024.08.16.608353

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