Burrill P H, Bernardini I, Kleinman H K, Kretchmer N
J Supramol Struct Cell Biochem. 1981;16(4):385-92. doi: 10.1002/jsscb.1981.380160409.
Rabbit intestinal epithelial cells, obtained after a limited hyaluronidase digestion, were incubated in medium with or without calf serum, on bacteriological plastic dishes. The dishes, either plain or coated with an air-dried type I collagen film, were pretreated with medium alone or eith medium containing purified laminin or purified fibronectin. Cells did not attach in significant numbers to untreated bacteriological plastic, even in the presence of serum. Cells did attach to collagen-coated dishes, and were judged viable on the basis of their incorporation of radiolabeled leucine into cell protein. Cell adhesion to the collagen substrate increased in proportion to the concentration of serum in the medium, with maximal attachment of 5% serum or greater. Pretreatment of plain or collagen-coated dishes with increasing amounts of fibronectin enhanced cell adhesion in a concentration-dependent manner. Either serum, or fibronectin-free serum in the medium enhanced cell attachment to substrates pretreated with either fibronectin or laminin. Thus, intestinal epithelial cells appear to possess surface receptors for both laminin and fibronectin. The evidence further suggests that calf serum may contain factors, other than fibronectin, capable of enhancing intestinal epithelial cell attachment to collagen substrates.
经有限的透明质酸酶消化后获得的兔肠上皮细胞,在有或无小牛血清的培养基中,于细菌学塑料培养皿上进行培养。这些培养皿,要么是普通的,要么涂有风干的I型胶原膜,先用单独的培养基或含有纯化层粘连蛋白或纯化纤连蛋白的培养基进行预处理。即使在有血清的情况下,细胞也不会大量附着于未处理的细菌学塑料上。细胞确实会附着于胶原包被的培养皿上,并根据它们将放射性标记的亮氨酸掺入细胞蛋白的情况判断为存活。细胞对胶原底物的黏附与培养基中血清的浓度成正比,血清浓度为5%或更高时黏附达到最大值。用越来越多的纤连蛋白预处理普通或胶原包被的培养皿,以浓度依赖的方式增强了细胞黏附。培养基中的血清或无纤连蛋白的血清均可增强细胞对用纤连蛋白或层粘连蛋白预处理的底物的附着。因此,肠上皮细胞似乎同时拥有层粘连蛋白和纤连蛋白的表面受体。证据还进一步表明,小牛血清可能含有除纤连蛋白之外的、能够增强肠上皮细胞对胶原底物附着的因子。