Genetically engineered elastin-like polymer as a substratum to culture cells from the ocular surface.
OBJECTIVE
To investigate epithelial cell adhesion and proliferation on a newly developed elastin-like polymer (ELP) that mimics the functional characteristics of extracellular matrices.


METHODS
A genetically engineered ELP with cell attachment sequences was adsorbed onto glass coverslips as 1, 2, or 3 molecular films. Conjunctival epithelial cells from a human cell line and human skin fibroblast cells (as controls) were plated onto coverslips with three different substrata: plain glass, Thermanox, and ELP-coated. Cells (10(4)) were plated after EDTA- or trypsin-based detachment. To test adhesion, epithelial and fibroblast cells were incubated for 4 hr, stained with hematoxylin, and counted. To study proliferation, Ki-67-positive epithelial cells were counted after 1, 3, and 5 days in culture. Immunostaining for conjunctival and adhesion markers was performed.


RESULTS
Epithelial cell, but not fibroblast, adhesion on ELP was significantly enhanced compared to that of control substrata. Epithelial cells detached with EDTA alone adhered significantly better than those detached with trypsin. By day 5, epithelial cell proliferation on ELP was significantly greater than that on plain glass. Epithelial cells grown on ELP expressed conjunctival and adhesion markers.


CONCLUSIONS
The recombinant ELP resembling the ocular surface extracellular matrix was a suitable substratum to sustain epithelial cell attachment and growth. This type of polymer may be suitable for tissue engineering to restore vision by reconstructing the ocular surface.
Martínez-Osorio
Hernán
H
Ocular Surface Group, Institute for Applied Ophthalmobiology (IOBA), University of Valladolid, Valladolid, Spain.
Juárez-Campo
Mónica
M
Diebold
Yolanda
Y
Girotti
Alessandra
A
Alonso
Matilde
M
Arias
Francisco Javier
FJ
Rodríguez-Cabello
José C
JC
García-Vázquez
Carmen
C
Calonge
Margarita
M
D001704:Biopolymers; D002448:Cell Adhesion / Q000502:physiology; D002452:Cell Count; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D003228:Conjunctiva / Q000166:cytology*; D004549:Elastin / Q000235:genetics*; D004847:Epithelial Cells / Q000166:cytology; D005109:Extracellular Matrix; D005347:Fibroblasts / Q000166:cytology; D005818:Genetic Engineering; D006801:Humans; D012867:Skin / Q000166:cytology
D001704:Biopolymers; D004549:Elastin