Isolation and characterization of Chinese hamster ovary cell variants defective in adhesion to fibronectin-coated collagen.
Isolation and characterization of Chinese hamster ovary cell variants defective in adhesion to fibronectin-coated collagen.
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DOI：
10.1083/jcb.87.3.755
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发表时间：
1980-12
影响因子：
7.8
通讯作者：
Juliano, R L
中科院分区：
文献类型：
--
作者：
Harper, P A;Juliano, R L
选择中国仓鼠卵巢（CHO）细胞的变体克隆，以降低对血清包被的组织培养板的粘附。这些克隆还显示出减少粘附到由包被有牛血清或纤连蛋白（冷不溶性球蛋白）的胶原蛋白层组成的基质。野生型（WT）和粘附变体（ADv）细胞在悬浮培养中以相当的速率生长，但粘附变体不能在单层培养中生长，因为它们不能附着在基质上。在这些细胞中的粘附缺陷没有得到纠正，通过提高二价阳离子或血清的浓度，以10倍以上的水平比那些通常用于细胞培养。然而，WT和ADv克隆可以粘附，传播，并获得一个正常的CHO形态上的基板包被刀豆球蛋白A或聚-L-赖氨酸。此外，粘附变体可以附着到涂覆有来源于人二倍体成纤维细胞或WT CHO细胞单层的“足垫”材料（附着于基质的材料）的基质上。这些观察结果表明，变异克隆可能具有细胞表面缺陷，这阻止它们利用外源性纤连蛋白作为促粘附配体;然而，变异体似乎具有正常的细胞骨架和代谢能力，使它们能够附着和扩散在涂覆有替代配体的基质上。这些变体在研究细胞粘附的分子基础方面应该是非常有用的。
Variant clones of Chinese hamster ovary (CHO) cells were selected for reduced adhesion to serum-coated tissue culture plates. These clones also displayed reduced adhesion to substrata composed of collagen layers coated with bovine serum or with fibronectin (cold-insoluble globulin). Wild-type (WT) and adhesion variant (ADv) cells grew at comparable rates in suspension culture, but the adhesion variants could not be grown in monolayer culture because of their inability to attach to the substratum. The adhesion deficit in these cells was not corrected by raising the concentration of divalent cations or of serum to levels 10-fold greater than those normally utilized in cell culture. However, both WT and ADv clones could adhere, spread, and attain a normal CHO morphology on substrata coated with concanavalin A or poly-L- lysine. In addition, the adhesion variants could attach to substrata coated with "footpad" material (substratum-attached material) derived from monolayers of human diploid fibroblasts or WT CHO cells. These observations suggest that the variant clones may have a cell surface defect that prevents them from utilizing exogeneous fibronectin as an adhesion-promoting ligand; however the variants seem to have normal cytoskeletal and metabolic capacities that allow them to attach and spread on substrata coated with alternative ligands. These variants should be extremely useful in studying the molecular basis of cell adhesion.