Garbi C, Nitsch L, Wollman S H
Exp Cell Res. 1984 Apr;151(2):458-65. doi: 10.1016/0014-4827(84)90395-1.
Separated thyroid follicles are stable in suspension culture in Coon's modified Ham's F12 medium containing 0.5% calf serum. They resemble follicles in vivo except for the absence of a basal lamina. However, the epithelial cells reverse polarity and the follicles invert when the serum concentration is raised to 5%. A number of substances, especially components of extracellular matrix, were added to the medium to ascertain if they could stabilize the follicles against inversion in 5% serum. Cellular and plasma fibronectin, gelatin, heat-denatured collagen, methylcellulose and laminin did not stabilize. The addition to the medium of as little as 50 micrograms/ml of acid-soluble collagen prepared from calf skin or rat tail tendons resulted in the formation of small clouds of gel. Follicles embedded within the gel were stabilized. Follicles in the same dish but not embedded in the gel inverted. Stabilization was not specific for collagen, since follicles embedded in a plasma clot were also stabilized. A gel was not sufficient for stabilization, since embedding in an agarose gel did not stabilize. Ultrastructural studies indicate that adherence to a limited number of gelled fibers of collagen covering only a small fraction of the basal plasma membrane may be sufficient to stabilize and that a basal lamina formed in the presence of laminin but without added collagen does not stabilize.
分离的甲状腺滤泡在含有0.5%小牛血清的Coon改良Ham's F12培养基中悬浮培养时是稳定的。除了没有基膜外,它们类似于体内的滤泡。然而,当血清浓度提高到5%时,上皮细胞极性反转,滤泡倒置。向培养基中添加了许多物质,特别是细胞外基质成分,以确定它们是否能使滤泡在5%血清中稳定而不倒置。细胞和血浆纤连蛋白、明胶、热变性胶原蛋白、甲基纤维素和层粘连蛋白都不能使其稳定。向培养基中添加低至50微克/毫升的从小牛皮或大鼠尾腱制备的酸溶性胶原蛋白会导致形成小凝胶云。包埋在凝胶中的滤泡得以稳定。同一培养皿中未包埋在凝胶中的滤泡则会倒置。稳定作用并非胶原蛋白所特有,因为包埋在血浆凝块中的滤泡也能稳定。凝胶不足以实现稳定,因为包埋在琼脂糖凝胶中并不能使其稳定。超微结构研究表明,仅附着于覆盖基底质膜一小部分的有限数量的胶凝胶原纤维可能足以实现稳定,并且在有层粘连蛋白但未添加胶原蛋白的情况下形成的基膜并不能实现稳定。