Mass Spectrometry Reveals α-2-HS-Glycoprotein as a Key Early Extracellular Matrix Protein for Conjunctival Cells

Makuloluwa, Aruni K, Stewart, Rosalind MK, Kaye, Stephen B ORCID: 0000-0003-0390-0592, Williams, Rachel L ORCID: 0000-0002-1954-0256 and Hamill, Kevin J ORCID: 0000-0002-7852-1944 (2020) Mass Spectrometry Reveals α-2-HS-Glycoprotein as a Key Early Extracellular Matrix Protein for Conjunctival Cells INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 61 (3). 44-. ISSN 0146-0404, 1552-5783

Text i1552-5783-276-1-28201.pdf - Published version Download (1MB) | Preview

Abstract

PURPOSE. To determine the composition of extracellular matrix (ECM) proteins secreted by a conjunctival epithelial cell line and to identify components that aid conjunctival epithelial cell culture. METHODS. Human conjunctival epithelial cell line (HCjE-Gi) cells were cultured in serum-free media and their ECM isolated using ammonium hydroxide. Growth characteristics were evaluated for fresh HCjE-Gi cells plated onto ECMs obtained from 3- to 28-day cell cultures. Mass spectrometry was used to characterize the ECM composition over 42 culture days. Cell adhesion and growth on pre-adsorbed fibronectin and α-2-HSglycoprotein (α-2-HS-GP) were investigated. RESULTS. Day 3 ECM provided the best substrate for cell growth compared to ECM obtained from 5- to 28-day cell cultures. Mass spectrometry identified a predominantly laminin 332 matrix throughout the time course, with progressive changes to matrix composition over time: proportional decreases in matrix-bound growth factors and increases in proteases. Fibronectin and α-2-HS-GP were 5- and 200-fold enriched as a proportion of the early ECM relative to the late ECM, respectively. Experiments on these proteins in isolation demonstrated that fibronectin supported rapid cell adhesion, whereas fibronectin and α-2-HS-GP both supported enhanced cell growth compared to tissue culture polystyrene. CONCLUSIONS. These data reveal α-2-HS-GP as a candidate protein to enhance the growth of conjunctival epithelial cells and raise the possibility of exploiting these findings for targeted improvement to synthetic tissue engineered conjunctival substrates.

Item Type:	Article
Uncontrolled Keywords:	ocular surface, conjunctiva, extracellular matrix, tissue engineering, mass spectrometry
Depositing User:	Symplectic Admin
Date Deposited:	17 Feb 2020 10:23
Last Modified:	16 Jun 2026 09:22
DOI:	10.1167/iovs.61.3.44
Related Websites:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3075032
Disclaimer:	The University of Liverpool is not responsible for content contained on other websites from links within repository metadata. Please contact us if you notice anything that appears incorrect or inappropriate.