Abstract
Cells are highly dynamic and adopt variable shapes and sizes. These variations are biologically important but challenging to investigate in a spatiotemporally controlled manner. Micropatterning, confining cells on microfabricated substrates with defined geometries and molecular compositions, is a powerful tool for controlling cell shape and interactions. However, conventional binary micropatterns are static and fail to address dynamic changes in cell polarity, spreading, and migration. Here, a method for dynamic micropatterning is reported, where the non-adhesive surface surrounding adhesive micropatterns is rapidly converted to support specific cell-matrix interactions while allowing simultaneous imaging of the cells. The technique is based on ultraviolet photopatterning of biotinylated polyethylene glycol-grafted poly-L-lysine, and it is simple, inexpensive, and compatible with a wide range of streptavidin-conjugated ligands. Experiments using biotinylation-based dynamic micropatterns reveal that distinct extracellular matrix ligands and bivalent integrin-clustering antibodies support different degrees of front-rear polarity in human glioblastoma cells, which correlates to altered directionality and persistence upon release and migration on fibronectin. Unexpectedly, however, neither an asymmetric cell shape nor centrosome orientation can fully predict the future direction of migration. Taken together, biotinylation-based dynamic micropatterns allow easily accessible and highly customizable control over cell morphology and motility.
| Original language | English |
|---|---|
| Article number | 2300719 |
| Number of pages | 17 |
| Journal | Small Methods |
| Volume | 8 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jan 2024 |
| MoE publication type | A1 Journal article-refereed |
Funding
The authors thank Petra Laasola and Jenni Siivonen for their expert technical assistance, Guillaume Jacquemet for help with microscopy, and the Ivaska Lab members for insightful feedback and discussion. The Cell Imaging and Cytometry Core facility (Turku Bioscience, University of Turku, Åbo Akademi University, and Biocenter Finland), and Turku Bioimaging are acknowledged for services, instrumentation, and expertise. This work was supported by the Finnish Cancer Institute (K. Albin Johansson Professorship to J.I.); Research Council of Finland research projects (grant no. 325464 to J.I.; 25700, 296684 and 307313 to P.K.M.) and Centre of Excellence program (grant no. 346131 to J.I.); the Cancer Foundation Finland (to J.I.); the Sigrid Jusélius Foundation (to J.I. and P.K.M.); the Jane and Aatos Erkko Foundation (to J.I.); the Research Council of Finland InFLAMES Flagship Programme (grant no. 337530); University of Turku Doctoral Programme in Molecular Life Sciences (to A.I.) and Molecular Medicine (to S.H-P.); the Finnish Cultural Foundation (to A.I. and S.H-P.); the Orion Research Foundation (to A.I.); and the K. Albin Johansson's Foundation (to A.I.). The authors thank Petra Laasola and Jenni Siivonen for their expert technical assistance, Guillaume Jacquemet for help with microscopy, and the Ivaska Lab members for insightful feedback and discussion. The Cell Imaging and Cytometry Core facility (Turku Bioscience, University of Turku, Åbo Akademi University, and Biocenter Finland), and Turku Bioimaging are acknowledged for services, instrumentation, and expertise. This work was supported by the Finnish Cancer Institute (K. Albin Johansson Professorship to J.I.); Research Council of Finland research projects (grant no. 325464 to J.I.; 25700, 296684 and 307313 to P.K.M.) and Centre of Excellence program (grant no. 346131 to J.I.); the Cancer Foundation Finland (to J.I.); the Sigrid Jusélius Foundation (to J.I. and P.K.M.); the Jane and Aatos Erkko Foundation (to J.I.); the Research Council of Finland InFLAMES Flagship Programme (grant no. 337530); University of Turku Doctoral Programme in Molecular Life Sciences (to A.I.) and Molecular Medicine (to S.H‐P.); the Finnish Cultural Foundation (to A.I. and S.H‐P.); the Orion Research Foundation (to A.I.); and the K. Albin Johansson's Foundation (to A.I.).
Keywords
- cell migration
- cell polarity
- extracellular matrices
- micropatterns