Evaluation of Müller cell attachment to various functional amino-acid sequences

Abstract

Purpose : Collagen is an abundantly distributed extracellular matrix protein that is known to maintain the structural integrity of the organs and tissues in mammals, and it not only plays a physical function as a structural protein, but also has various biological functions that regulate cell adhesion, migration, and differentiation. In this study, we investigated the adhesion activity of cells in the human Müller cell line MIO-M1 with certain amino acid sequences displayed on the collagen triple helices, which can control cell adhesion, proliferation, and survival by arbitrarily incorporating collagen-derived functional amino-acid sequences.

Methods : The wells of 96-well plates were coated with a peptide polymer containing four kinds of functional amino-acid sequences (GFOGER, GLOGEN, GVXGFO, and KGHRGF) at the concentrations of 1, 3, 10, 30, and 100%, and MIO-M1 cell-line cells (5 x 104 cells/well) were then seeded. After incubation at 37°C for 1 hour, the wells were washed twice with phosphate buffered saline (PBS), and then incubated again at 37°C for 1 hour. The Cell Counting Kit-8^® (CCK-8; Dojindo Laboratories) viability assay was then used to evaluate cell adhesion. Wells without coating were used as controls.

Results : In the GFOGER group, GLOGEN group, and GVXGFO group, cell viability was higher in a concentration-dependent manner. In the GFOGER group and the GLOGEN group, the cell viability was significantly higher in the concentration of 100% compared to the control, 1, 3, and 10%, respectively (p<0.01, paired t test), while there were no significant differences between 100% and 30%. In the GVXGFO group and the KGHRGF group, the concentration of 100% showed significantly higher cell viability than the control and other concentrations (p<0.01, paired t test).

Conclusions : The findings in this study showed that collagen-derived functional amino-acid sequences improved the adhesion activity of MIO-M1 cell-line cells.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.