Research ArticleNon-tumor mast cells cultured in vitro on a honeycomb-like structured film proliferate with multinucleated formation
Graphical Abstract
Various steps of multinuclear NCL-2 cell (non-tumor mast cell) proliferation on the 3-μm honeycomb-like structured polystyrene film (HCF). A cell adhering to the 3-μm HCF (A), cell ballooning and division along the structure of the hole (B, C), a splitting cell migrating down into the hole of the film (D, E), and individual cells along the holes (F). The images of each stage were not obtained from the same cells. All scale bars are 5 μm.
Section snippets
Preparation of honeycomb-like films (HCFs)
Porous HCFs of polystyrene were fabricated using water droplets as templates for regular pores. In brief, polystyrene, didecylacrylamide, and w-carboxyhexylacrylamide (CAP) (10:1 wt%) were dissolved in chloroform at a concentration of 5 g/L, and the polymer mixture was layered on cover glasses in a humid environment. The pore size was regulated by changing the casting conditions (temperature, humidity, gas flow rate, casting volume, and casting concentration) of the polymer solution.8, 9
NCL-2 cell line
Cell
NCL-2 cell proliferation on polystyrene films
NCL-2 cells are essentially non-adherent cells, but some do attach to the ECM. At 1 week after seeding at a density of 1 × 105 cells/ml, the total number of NCL-2 cells (adherent plus floating cells) gradually increased with increasing HCF hole size. After 1 week, the NCL-2 cells reached 10.72 × 105 cells/ml on the flat film, 15.36 × 105 cells/ml on the 3-μm HCF, 15.39 × 105 cells/ml on the 5-μm HCF, and 18.53 × 105 cells/ml on the 10-μm HCF. The number of NCL-2 cells adhering to the film was 0.72 × 105
Discussion
In mammalian cells, nuclear division is usually followed by cytoplasmic division. However, megakaryocytes, which produce blood platelets, and some hepatocytes and heart muscle cells become multinucleated.11 Because mast cells are derived from bone marrow hematopoietic cells, they may have the same characteristic cell division pattern as megakaryocytes. We propose that HCFs exhibiting self-organization and a suitable three-dimensional (3D) pore size could provide a physical stimulus for mast
Acknowledgments
The authors would like to thank FUJIFILM Corporation (Ltd), Kanagawa, Japan, for supplying the HCFs.
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Conflict of Interest: The authors declare no competing financial interests.