Cell type-specific adaptation of cellular and nuclear volume in micro-engineered 3D environments.
Bio-functionalized three-dimensional (3D) structures fabricated by direct laser writing (DLW) are structurally and mechanically well-defined and ideal for systematically investigating the influence of three-dimensionality and substrate stiffness on cell behavior. Here, we show that different fibroblast-like and epithelial cell lines maintain normal proliferation rates and form functional cell-matrix contacts in DLW-fabricated 3D scaffolds of different mechanics and geometry. Furthermore, the molecular composition of cell-matrix contacts forming in these 3D micro-environments and under conventional 2D culture conditions is identical, based on the analysis of several marker proteins (paxillin, phospho-paxillin, phospho-focal adhesion kinase, vinculin, β1-integrin). However, fibroblast-like and epithelial cells differ markedly in the way they adapt their total cell and nuclear volumes in 3D environments. While fibroblast-like cell lines display significantly increased cell and nuclear volumes in 3D substrates compared to 2D substrates, epithelial cells retain similar cell and nuclear volumes in 2D and 3D environments. Despite differential cell volume regulation between fibroblasts and epithelial cells in 3D environments, the nucleus-to-cell (N/C) volume ratios remain constant for all cell types and culture conditions. Thus, changes in cell and nuclear volume during the transition from 2D to 3D environments are strongly cell type-dependent, but independent of scaffold stiffness, while cells maintain the N/C ratio regardless of culture conditions.
Karlsruhe Institute of Technology (KIT), Zoological Institute, Department of Cell- and Neurobiology, Haid-und-Neu-Straße 9, D-76131 Karlsruhe, Germany.
Alexandra M
AM
Greiner
Karlsruhe Institute of Technology (KIT), Zoological Institute, Department of Cell- and Neurobiology, Haid-und-Neu-Straße 9, D-76131 Karlsruhe, Germany.
Franziska
F
Klein
Karlsruhe Institute of Technology (KIT), DFG-Center for Functional Nanostructures (CFN), Wolfgang-Gaede-Straße 1a, D-76131 Karlsruhe, Germany.
Tetyana
T
Gudzenko
Karlsruhe Institute of Technology (KIT), Zoological Institute, Department of Cell- and Neurobiology, Haid-und-Neu-Straße 9, D-76131 Karlsruhe, Germany.
Benjamin
B
Richter
Karlsruhe Institute of Technology (KIT), Zoological Institute, Department of Cell- and Neurobiology, Haid-und-Neu-Straße 9, D-76131 Karlsruhe, Germany.
Thomas
T
Striebel
Karlsruhe Institute of Technology (KIT), Zoological Institute, Department of Cell- and Neurobiology, Haid-und-Neu-Straße 9, D-76131 Karlsruhe, Germany.
Bayu G
BG
Wundari
Karlsruhe Institute of Technology (KIT), Institute of Functional Interfaces (IFG), 76344 Eggenstein-Leopoldshafen, Germany.
Tatjana J
TJ
Autenrieth
Karlsruhe Institute of Technology (KIT), DFG-Center for Functional Nanostructures (CFN), Wolfgang-Gaede-Straße 1a, D-76131 Karlsruhe, Germany; Karlsruhe Institute of Technology (KIT), Institute for Applied Physics, Wolfgang-Gaede-Straße 1, D-76131 Karlsruhe, Germany.
Martin
M
Wegener
Karlsruhe Institute of Technology (KIT), DFG-Center for Functional Nanostructures (CFN), Wolfgang-Gaede-Straße 1a, D-76131 Karlsruhe, Germany. Electronic address: clemens.franz@kit.edu.
Clemens M
CM
Franz
Karlsruhe Institute of Technology (KIT), Zoological Institute, Department of Cell- and Neurobiology, Haid-und-Neu-Straße 9, D-76131 Karlsruhe, Germany; Karlsruhe Institute of Technology (KIT), DFG-Center for Functional Nanostructures (CFN), Wolfgang-Gaede-Straße 1a, D-76131 Karlsruhe, Germany; Karlsruhe Institute of Technology (KIT), Institute of Functional Interfaces (IFG), 76344 Eggenstein-Leopoldshafen, Germany. Electronic address: bastmeyer@kit.edu.
Martin
M
Bastmeyer
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002460:Cell Line; D049109:Cell Proliferation; D048429:Cell Size; D004847:Epithelial Cells / Q000166:cytology*; D004867:Equipment Design; D005109:Extracellular Matrix / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology*; D007834:Lasers; D051379:Mice; D051381:Rats; D054457:Tissue Scaffolds / Q000737:chemistry*
3D cell culture substrates; Cell volume; Cell-matrix adhesions; Direct laser writing; Nuclear volume