Effect of culture substrate and culture conditions on lens epithelial cell proliferation and alpha-smooth muscle actin expression.
The most common complication following cataract surgery is posterior capsule opacification. This results from migration, proliferation and transdifferentiation of residual lens epithelial cells (LECs). We studied the effect of several culture substrates and culture conditions on LEC proliferation and alpha-smooth muscle actin (alpha-SMA) expression. We used primary and secondary cultures of porcine LECs cultivated on collagen I, collagen IV, microscopic glass slides, and uncoated plastic dishes. We studied the cell proliferation and expression of alpha-SMA and alpha-, beta-, and gamma-crystallins. The effect of the medium exchange protocol was studied using the TOTL-86 rabbit epithelial lens cell line. There was no difference in growth characteristics of primary cultures on different substrates. In secondary cultures, LECs adhered better to collagen-coated surfaces. The culture substrate influenced LEC proliferation and alpha-SMA expression. The proliferation was greater when the medium was changed than when extra medium was added on the 4th day. The cells did not synthesize alpha-, beta- or gamma-crystallin. The culture substrate influences the adhesion ability, proliferation and alpha-SMA expression in lens epithelial cells. In addition, it is necessary to consider the effects of the medium exchange protocol, serum supplementation, cell density and other cell culture conditions in lens epithelial cell experiments.
Mahelková
G
G
2nd Faculty of Medicine, Institute of Physiology, Charles University in Prague, Prague, Czech Republic.
Bacáková
L
L
Korynta
J
J
Vajner
L
L
Vytásek
R
R
D000199:Actins / Q000378:metabolism*; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D007150:Immunohistochemistry; D007908:Lens, Crystalline / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects*; D011817:Rabbits; D013552:Swine; D038201:alpha-Crystallins / Q000378:metabolism; D038204:beta-Crystallins / Q000378:metabolism; D038222:gamma-Crystallins / Q000378:metabolism
D000199:Actins; D003470:Culture Media; D038201:alpha-Crystallins; D038204:beta-Crystallins; D038222:gamma-Crystallins; D003094:Collagen