Gosselin F, Magloire H, Joffre A, Portier M M
Laboratoire de Biochimie Cellulaire, Collège de France, Paris.
Arch Oral Biol. 1990;35 Suppl:217S-221S. doi: 10.1016/0003-9969(90)90162-4.
Cytokeratins are considered to be molecular markers for different types of epithelial differentiation. They were used to investigate the precise differentiation stage of gingival epithelium, reconstituted in vitro, following two different culture procedures. Human trypsin-dissociated gingival keratinocytes were seeded either on a feeder layer of irradiated mouse 3T3 fibroblasts or on a connective tissue equivalent (lattice) made up of human fibroblasts in a collagen gel. The cytokeratins were extracted and analysed by two-dimensional gel electrophoresis. Although both methods showed on histological sections that cultured gingival keratinocytes formed a multilayered non-keratinizing epithelium, the cytokeratins patterns showed great differences. The gingival epithelium-like structure reconstituted on 3T3 feeder layer expressed some cytokeratins characteristic of the in situ gingival epithelium (K 5, 6, 14, 16, 17) and some which do not exist in the normal tissue (K 8, 18, 19, traces of K 13 and K 15) and are specific for embryonic, simple and tumour epithelia. However, the gingival epithelium reconstituted on connective tissue equivalent expressed all the cytokeratins present in the normal tissue (K 5, 6, 14, 16, 17), except those specific for terminal differentiation (K 1, 2, and 10/11). These findings suggest that the culture of gingival keratinocytes on connective tissue equivalents allows them to reproduce physiological stages of differentiation.
细胞角蛋白被认为是不同类型上皮分化的分子标志物。它们被用于研究体外重建的牙龈上皮在两种不同培养程序后的精确分化阶段。将人胰蛋白酶解离的牙龈角质形成细胞接种在经辐照的小鼠3T3成纤维细胞饲养层上，或接种在由人成纤维细胞在胶原凝胶中构成的结缔组织替代物（网格）上。通过二维凝胶电泳提取并分析细胞角蛋白。尽管两种方法在组织学切片上均显示培养的牙龈角质形成细胞形成了多层非角化上皮，但细胞角蛋白模式显示出很大差异。在3T3饲养层上重建的牙龈上皮样结构表达了一些原位牙龈上皮特有的细胞角蛋白（K5、6、14、16、17）以及一些正常组织中不存在的细胞角蛋白（K8、18、19、微量的K13和K15），这些细胞角蛋白对胚胎、单层和肿瘤上皮具有特异性。然而，在结缔组织替代物上重建的牙龈上皮表达了正常组织中存在的所有细胞角蛋白（K5、6、14、16、17），但不包括终末分化特有的那些（K1、2和10/11）。这些发现表明，在结缔组织替代物上培养牙龈角质形成细胞能使其重现生理分化阶段。
