Effect of cell growth supporting surface on phagocytosis

Girish K Srivastava 1, 2, David Rodriguez-Crespo 1, J. Carlos Pastor 1, 2
1. Instituto Universitario de Oftalmobiologia Aplicada (IOBA), Universidad de Valladolid, Valladolid, Spain.
2. Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Spain.
Abstract

Phagocytosis is involved in several functions including nutrient uptake, immune response, inflammation, tissue homeostasis, cellular apoptotic bodies and debris elimination. Its dysfunction has impacts on normal functioning of an organism. It is involved in several diseases including the AMD pathogenesis, and treatment approaches. Mostly an initial proof of concept is tested in a laboratory conditions. Cell and tissue cultures need proper surfaces to grow and maintain them in laboratory conditions. The study aims to evaluate effect of two surfaces; tissue culture plate polystyrene and glass coverslip, on phagocytosis performed by RPE cells growing on each surface. Fresh RPE cells and ARPE-19 cell line were grown in corresponding cell culture conditions. Phagocytosis in fresh RPE cell cultures was detected and recorded. ARPE-19 cells were incubated for 4 and 24 hours with 0.2 and 1 μl red fluorescent latex beads, and then images were taken to measure the average numbers of cells, cells performing no phagocytosis and phagocytosis and engulfed red fluorescent latex beads. Results showed that RPE cell membrane was forming blebs releasing black dots like particles as cell debris in cell culture medium. Subsequently cytoplasmic membrane extensions were formed which had engulfed black dots like particles. ARPE-19 cells are a good cellular model for performing initial in vitro test in comparison to fresh RPE cells. There was no significant difference between numbers of ARPE-19 cells growing on polystyrene and glass surface performing latex bead phagocytosis. Both had engulfed red fluorescent latex beads; however, numbers of engulfed beads was significantly higher for glass surface than polystyrene surface. Results support the hypothesis that surface of cell culture plates affects the cellular phagocytosis mechanism which play a critical role in the functions in many cases such as in vivo retina. These findings are very crucial when it comes to transfer a treatment approach based on phagocytosis to the clinics and that the materials and the conditions of the experiments can have very important consequences.    
Keywords: Retinal pigment epithelial cells, phagocytosis, cell growth supporting surface, latex beads  

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Publication
Ms. Ref. No.: A0201006     Reviewer    Author
Submitted: 26/07/2018
Accepted:15/09/2018
Published Online: 15/01/2020

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