Reichard-Brown J L, Akeson R
Dev Biol. 1983 Apr;96(2):304-16. doi: 10.1016/0012-1606(83)90167-7.
The differential adhesion of cultured mammalian clonal cell lines to components of the extracellular matrix was examined by kinetic adhesion and long-term growth assays. Uniform artificial matrices were prepared by air drying collagen Type I solution (C) onto a microtiter well and then air drying a solution containing a single glycosaminoglycan (GAG): hyaluronic acid (HA), chondroitin sulfate-4 (CHS-4), or chondroitin sulfate-6 (CHS-6). The adhesion of [3H]thymidine-prelabeled cells suspended in fibronectin (FN) depleted medium was measured at 2 and 6 hr. Neuroblastoma (N18, Lan 1) and melanoma (B16, G361, S91) cell lines exhibited a significantly greater percentage of cells adhering to one or more C-GAG matrices compared with C matrices. Maximal adhesion at 2 hr was to C-HA. In contrast at 2 hr, two glial, two epithelial, and one fibroblastic cell line showed unchanged or significantly decreased binding to C-GAG compared with C matrices. Further experiments using a neuroblastoma (N18) and a glioma (C6) cell line indicated that the adhesion patterns were not altered either by the method of dissociation from the tissue culture dish, preincubation with exogenous GAG, or the addition of exogenous fibronectin. Assays of N18 and C6 adhesion to matrices made from a non-GAG polyanionic compound, polygalacturonic acid (PGA), did not yield the same adhesion patterns as C-HA matrices. Long-term growth studies of a neuroblastoma (N18) melanoma (S91), and glioma (C6) cell line on nonuniform matrices deliberately prepared with GAG-rich and GAG-poor regions complemented the observations from the kinetic adhesion assays. N18 and S91 cells did not grow on areas which did not contain GAG by toluidine blue staining. However, the C6 cells did not grow on areas which did strongly stain for GAG. A quantitative analysis of the long term growth of N18 and C6 cells substantiated these observations. All these data indicate that the cellular phenotype may be correlated with matrix adhesion. Neuroblastomas and melanomas have a greater affinity for GAG-containing matrices while glial, epithelial, and fibroblastic cells appear to have a greater or equal affinity for collagen matrices.
通过动力学黏附试验和长期生长试验，研究了培养的哺乳动物克隆细胞系对细胞外基质成分的差异黏附情况。通过将I型胶原溶液（C）在微量滴定板上空气干燥，然后将含有单一糖胺聚糖（GAG）的溶液空气干燥来制备均匀的人工基质，所述单一糖胺聚糖为透明质酸（HA）、硫酸软骨素-4（CHS-4）或硫酸软骨素-6（CHS-6）。在2小时和6小时时，测量悬浮在纤连蛋白（FN）缺失培养基中的[3H]胸腺嘧啶预标记细胞的黏附情况。与C基质相比，神经母细胞瘤（N18、Lan 1）和黑色素瘤（B16、G361、S91）细胞系黏附于一种或多种C-GAG基质的细胞百分比显著更高。2小时时最大黏附发生在C-HA上。相比之下在2小时时，两种神经胶质细胞系、两种上皮细胞系和一种成纤维细胞系与C基质相比，对C-GAG的结合未改变或显著降低。使用神经母细胞瘤（N18）和胶质瘤（C6）细胞系进行的进一步实验表明，黏附模式不会因从组织培养皿解离的方法、与外源性GAG预孵育或添加外源性纤连蛋白而改变。对由非GAG多阴离子化合物聚半乳糖醛酸（PGA）制成的基质进行的N18和C6黏附试验，未产生与C-HA基质相同的黏附模式。对故意制备有富含GAG和缺乏GAG区域的非均匀基质上的神经母细胞瘤（N18）、黑色素瘤（S91）和胶质瘤（C6）细胞系进行的长期生长研究，补充了动力学黏附试验的观察结果。通过甲苯胺蓝染色，N18和S91细胞在不含GAG的区域不生长。然而，C6细胞在对GAG染色强烈的区域不生长。对N18和C
