Regensburg 2016 – scientific programme
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BP: Fachverband Biologische Physik
BP 52: Posters - Multi-Cellular Systems
BP 52.3: Poster
Wednesday, March 9, 2016, 17:00–19:00, Poster C
Quantifying Cell Volume and Proliferation in MDCK II Model Tissues — •Simone Gehrer1, Damir Vurnek1, Sara Kaliman1, Florian Rehfeldt2, Diana Dudziak3, and Ana-Sunčana Smith1, 4 — 1PULS Group, Institute of Theoretical Physics I, FAU Erlangen — 23rd Institute of Physics/Biophysics, GAU Göttingen — 3University Hospital Erlangen — 4Division of Physical Chemistry, IRB Zagreb
Multicellular migration of 2D cell sheets is essential for wound healing and organ development. In-vitro model systems show that cell volume and proliferation are key players in active epithelial rearrangement and tissue expansion. Although studied extensively, precise temporal and spatial quantitative analysis of proliferation still remains a challenge.
To reveal the dependence of cell volume and proliferation on different monolayer compartments we droplet seeded MDCK II cells on collagen I coated glass. Grown from subconfluency to fully compartmentalized clusters they were examined on days 1, 2, 4 and 6. Volume studies were carried out on confocal images of actin and DAPI stained samples. As a part of the testing procedures the analysis was repeated on the stably transfected cell line with GFP histones generated in our lab. To visualize proliferation the monolayers were preincubated with EdU, stained, and imaged with fluorescence microscopy. Cells were counted individually with a home-written Matlab routine and the radial distribution of cell proliferation was obtained. For proliferation an inverse dependence on density was confirmed. Surprisingly, we found a direct correlation between the proliferation and the age of the tissues.