Fröhlich E, Wahl R, Reutter K
Department of Anatomy, Eberhard-Karls University of Tübingen, Germany.
Histochem J. 1995 Aug;27(8):602-8.
Porcine thyroid cells isolated by dispase treatment were cultured in either (a) Matrigel, (b) agarose with the addition of different combinations of basic fibroblast growth factor and laminin, or (c) on agarose-coated dishes. The formation of follicles and the presence of a basal lamina was investigated by routine electron microscopy of Araldite-embedded material and by light and electron microscopical immunocytochemical detection of the basal lamina components, laminin and collagen type IV. After 10 days of culture in Matrigel or agarose, a basal lamina-like structure surrounded most follicles. Follicles of cells growing in agarose and overlaid with a medium containing thyrotropin and overlaid with a medium containing thyrotropin and fibroblast growth factor showed a fluorescent band at the basal side of the follicles after immunocytochemical staining with anti-laminin and anti-collagen antibodies. Routine electron microscopy showed that a basal lamina-like structure lined the outside of the follicle. This structure could be subdivided into a lamina lucida and a lamina densa. Electron microscopical immunogold labelling revealed that immunologically detectable laminin was confined to the lamina densa. These findings suggest that even in the absence of basal lamina components in the culture medium, thyroid cells are able to form follicles with a regular basal lamina when they are cultured in a three-dimensional environment.
通过胰蛋白酶处理分离得到的猪甲状腺细胞,分别培养于以下环境中:(a)基质胶;(b)添加了不同组合的碱性成纤维细胞生长因子和层粘连蛋白的琼脂糖;或(c)琼脂糖包被的培养皿。通过对包埋于环氧树脂中的材料进行常规电子显微镜检查,以及通过对基底膜成分层粘连蛋白和IV型胶原进行光镜和电镜免疫细胞化学检测,来研究滤泡的形成和基底膜的存在情况。在基质胶或琼脂糖中培养10天后,大多数滤泡被类似基底膜的结构所包围。在琼脂糖中生长且覆盖含有促甲状腺激素的培养基以及覆盖含有促甲状腺激素和成纤维细胞生长因子的培养基的细胞滤泡,在用抗层粘连蛋白和抗胶原抗体进行免疫细胞化学染色后,在滤泡基底侧显示出一条荧光带。常规电子显微镜检查显示,类似基底膜的结构排列在滤泡外部。该结构可细分为透明层和致密层。电子显微镜免疫金标记显示,免疫可检测到的层粘连蛋白局限于致密层。这些发现表明,即使在培养基中不存在基底膜成分的情况下,甲状腺细胞在三维环境中培养时仍能够形成具有规则基底膜的滤泡。