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培养底物和培养条件对晶状体上皮细胞增殖及α-平滑肌肌动蛋白表达的影响

Effect of culture substrate and culture conditions on lens epithelial cell proliferation and alpha-smooth muscle actin expression.

作者信息

Mahelková G, Bacáková L, Korynta J, Vajner L, Vytásek R

机构信息

2nd Faculty of Medicine, Institute of Physiology, Charles University in Prague, Prague, Czech Republic.

出版信息

Folia Biol (Praha). 2009;55(2):66-76.

Abstract

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.

摘要

白内障手术后最常见的并发症是后囊膜混浊。这是由残留晶状体上皮细胞(LECs)的迁移、增殖和转分化引起的。我们研究了几种培养底物和培养条件对LECs增殖及α-平滑肌肌动蛋白(α-SMA)表达的影响。我们使用了在I型胶原、IV型胶原、显微镜载玻片和未包被的塑料培养皿上培养的猪LECs原代和传代培养物。我们研究了细胞增殖以及α-SMA和α-、β-、γ-晶状体蛋白的表达。使用TOTL-86兔晶状体上皮细胞系研究了培养基更换方案的影响。不同底物上原代培养物的生长特性没有差异。在传代培养中,LECs在胶原包被的表面附着更好。培养底物影响LECs增殖和α-SMA表达。第4天更换培养基时的增殖比添加额外培养基时更大。细胞不合成α-、β-或γ-晶状体蛋白。培养底物影响晶状体上皮细胞的黏附能力、增殖和α-SMA表达。此外,在晶状体上皮细胞实验中,有必要考虑培养基更换方案、血清补充、细胞密度和其他细胞培养条件的影响。

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