In vitro evaluation of the interactions between human corneal endothelial cells and extracellular matrix proteins.
The corneal endothelium is the innermost cell layer of the cornea and rests on Descemet's membrane consisting of various extracellular matrix (ECM) proteins which can directly affect the cellular behaviors such as cell adhesion, proliferation, polarity, morphogenesis and function. The objective of this study was to investigate the interactions between the ECM environment and human corneal endothelial cells (HCECs), with the ultimate goal to improve cell proliferation and function in vitro. To evaluate the interaction of HCECs with ECM proteins, cells were seeded on ECM-coated tissue culture dishes, including collagen type I (COL I), collagen type IV (COL IV), fibronectin (FN), FNC coating mix (FNC) and laminin (LM). Cell adhesion and proliferation of HCECs on each substratum and expression of CEC markers were studied. The results showed that HCECs plated on the COL I, COL IV, FN and FNC-coated plates had enhanced cell adhesion initially; the number for COL I, COL IV, FN and FNC was significantly higher than the control (P < 0.05). In addition, cells grown on ECM protein-coated dishes showed more compact cellular morphology and CEC marker expression compared to cells seeded on uncoated dishes. Collectively, our results suggest that an adequate ECM protein combination can provide a long-term culture environment for HCECs for corneal endothelium transplantation.
Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
Jin San
JS
Choi
Eun Young
EY
Kim
Min Jeong
MJ
Kim
Matthew
M
Giegengack
Faraaz A
FA
Khan
Gilson
G
Khang
Shay
S
Soker
D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000235:genetics; D049109:Cell Proliferation; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D003094:Collagen / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004728:Endothelium, Corneal / Q000166:cytology* / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D016023:Integrins / Q000235:genetics; D007797:Laminin / Q000737:chemistry; D008422:Materials Testing; D046509:Tissue Culture Techniques
D015815:Cell Adhesion Molecules; D020099:Coated Materials, Biocompatible; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D016023:Integrins; D007797:Laminin; D003094:Collagen