Spheroid formation of human keratinocyte: Balancing between cell-substrate and cell-cell interaction
Issue title: Selected papers of the 39th Conference of the German Society for Clinical Microcirculation and Hemorheology, 6-7 November 2020, Hannover, Germany
Guest editors: B. Hiebl, A. Krüger-Genge and F. Jung
Affiliations: [a]
Institute of Biomaterial Science and Berlin-Brandenburg Center for Regenerative Therapies, Helmholtz-Zentrum Geesthacht, Teltow, Germany
| [b] Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany
| [c] Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany
Correspondence:
[*]
Corresponding authors: Prof. Dr. Nan Ma, E-mail: nan.ma@hzg.de and Prof. Dr. Andreas Lendlein, E-mail: andreas.lendlein@hzg.de.
Abstract: BACKGROUND:The formation of spheroids is tightly regulated by intrinsic cell-cell and cell-substrate interactions. OBJECTIVE:The chitosan (CS)-coating was applied to investigate the driven force directed the spheroid formation. METHODS:The effects of CS on cell functions were studied. Atomic force microscopy was employed to measure the cell- biomaterial interplay at single cell level. RESULTS:HaCaT cells shifted from their flattened sheet to a compact 3D spheroidal morphology when increasing CS-coating concentration. The proliferative capacity of HaCaT was preserved in the spheroid. The expression and activation of integrin β1 (ITGB1) were enhanced on CS modified surfaces, while the active to total ratio of ITGB1 was decreased. The adhesive force of a single HaCaT cell to the tissue culture plate (TCP) was 4.84±0.72 nN. It decreased on CS-coated surfaces as CS concentration increased, from 2.16±0.26 nN to 0.96±0.17 nN. The adhesive force between the single HaCaT cell to its neighbor cell increased as CS concentration increased, from 1.15±0.09 nN to 2.60±0.51 nN. CONCLUSIONS:Conclusively, the decreased cell- substrate adhesion was the main driven force in the spheroid formation. This finding might serve as a design criterion for biomaterials facilitating the formation of epithelial spheroids.
