Oharazawa H, Ibaraki N, Lin L R, Reddy V N
Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
Exp Eye Res. 1999 Dec;69(6):603-10. doi: 10.1006/exer.1999.0723.
Lens capsule consists of several kinds of extracellular matrix (ECM) which may play an important role in cell attachment, migration and proliferation of lens epithelial cells as a basement membrane. We have investigated the effects of ECM on cell attachment, proliferation and migration in a human lens epithelial (HLE) cell line. The HLE cell line, SRA 01/04, which was transfected with large T-antigen of SV40 was cultured in the absence of serum. Culture plates were coated with human type IV collagen, laminin or fibronectin. The number of cells were counted at 30-180 min and 3, 5 and 7 days of culture. The rate of BrdU incorporation was measured to study the cell proliferation. The cell migration was measured 1, 3, 5 and 7 days after seeding cells. Integrins, the receptors of ECM, were also detected using antibodies for the cell membrane antigens (CD49b, CD49c, CD49e) by an immunohistochemical method. Although less than 10% of cells attached to the non-coated plate and 50-60% of cells attached to the ECM-coated plates, there was no difference of cell attachment among each ECM used. The cell attachment was almost complete during the first 30 min of culture. Cell proliferation was not enhanced, but cell survival was aided by culture on the ECM components for up to 7 days. The area of cell attachment enlarged on the ECM-coated plates, whereas no migration was observed on the non-coated plate. These data indicate that ECM is the essential factor for cell attachment and increases migration of HLE cells.
晶状体囊膜由几种细胞外基质(ECM)组成,作为基底膜,其可能在晶状体上皮细胞的细胞附着、迁移和增殖中发挥重要作用。我们研究了ECM对人晶状体上皮(HLE)细胞系中细胞附着、增殖和迁移的影响。用SV40大T抗原转染的HLE细胞系SRA 01/04在无血清条件下培养。培养板用人类IV型胶原、层粘连蛋白或纤连蛋白包被。在培养30 - 180分钟以及3、5和7天时对细胞数量进行计数。测量BrdU掺入率以研究细胞增殖。在接种细胞后1、3、5和7天测量细胞迁移。还通过免疫组织化学方法使用针对细胞膜抗原(CD49b、CD49c、CD49e)的抗体检测ECM的受体整合素。尽管不到10%的细胞附着在未包被的培养板上,50 - 60%的细胞附着在ECM包被的培养板上,但所使用的每种ECM之间细胞附着没有差异。在培养的前30分钟内细胞附着几乎完成。细胞增殖未增强,但在ECM成分上培养长达7天有助于细胞存活。在ECM包被的培养板上细胞附着面积增大,而在未包被的培养板上未观察到迁移。这些数据表明ECM是细胞附着的关键因素,并增加HLE细胞的迁移。