Nicolaissen B, Haaskjold E, Fiskaadal H J, Vegge T
Department of Ophthalmology, Ulleval Hospital, Norway.
Acta Ophthalmol (Copenh). 1989 Aug;67(4):435-40. doi: 10.1111/j.1755-3768.1989.tb01629.x.
Retinal pigment epithelium from human eyes was maintained in organ culture, and DNA synthesis was visualized by autoradiography of cultures exposed to labelled thymidine. In Ham's F10 medium, labelling of cells was observed after 7 as well as after 14 days in vitro. At both stages the labelled cells were characteristically located adjacent to areas of epithelial damage. Cells remote from such areas did not incorporate thymidine, This pattern was observed in epithelia removed from the posterior pole as well as from the periphery of the fundus. Several types of routine media and media with human as well as foetal bovine serum (FBS) were found to support DNA synthesis in the cultured epithelium, labelled cells were not observed in epithelium maintained in Ham's F10 without serum, or with low serum concentration. Our study demonstrates, that DNA synthesis does take place in organ cultured human retinal pigment epithelium when the appropriate culture conditions are used. Our findings indicate that induction of DNA synthesis in the cells is not promoted by an indiscriminate effect of the medium but related to the presence of epithelial damage.
人眼视网膜色素上皮细胞被维持在器官培养中,通过对暴露于标记胸腺嘧啶核苷的培养物进行放射自显影来观察DNA合成。在Ham's F10培养基中,体外培养7天和14天后均观察到细胞标记。在这两个阶段,标记细胞的特征性位置是靠近上皮损伤区域。远离这些区域的细胞不掺入胸腺嘧啶核苷。这种模式在从后极以及眼底周边取出的上皮中均有观察到。发现几种常规培养基以及含人血清和胎牛血清(FBS)的培养基可支持培养的上皮细胞中的DNA合成,在无血清或血清浓度低的Ham's F10培养基中培养的上皮中未观察到标记细胞。我们的研究表明,当使用适当的培养条件时,器官培养的人视网膜色素上皮细胞中确实会发生DNA合成。我们的发现表明,细胞中DNA合成的诱导不是由培养基的无差别作用促进的,而是与上皮损伤的存在有关。