Curtis A S, Forrester J V, McInnes C, Lawrie F
J Cell Biol. 1983 Nov;97(5 Pt 1):1500-6. doi: 10.1083/jcb.97.5.1500.
The surface treatment of polystyrene, which is required to make polystyrene suitable for cell adhesion and spreading, was investigated. Examination of surfaces treated with sulfuric acid or various oxidizing agents using (a) x-ray photoelectron and attenuated total reflection spectroscopy and (b) measurement of surface carboxyl-, hydroxyl-, and sulfur-containing groups by various radiochemical methods showed that sulfuric acid produces an insignificant number of sulfonic acid groups on polystyrene. This technique together with various oxidation techniques that render surfaces suitable for cell culture generated high surface densities of hydroxyl groups. The importance of surface hydroxyl groups for the adhesion of baby hamster kidney cells or leukocytes was demonstrated by the inhibition of adhesion when these groups were blocked: blocking of carboxyl groups did not inhibit adhesion and may raise the adhesion of a surface. These results applied to cell adhesion in the presence and absence of serum. The relative unimportance of fibronectin for the adhesion and spreading of baby hamster kidney cells to hydroxyl-rich surfaces was concluded when cells spread on such surfaces after protein synthesis was inhibited with cycloheximide, fibronectin was removed by trypsinization, and trypsin activity was stopped with leupeptin.
为使聚苯乙烯适用于细胞黏附和铺展,对其表面处理进行了研究。使用(a)X射线光电子能谱和衰减全反射光谱,以及(b)通过各种放射化学方法测量表面含羧基、羟基和硫的基团,对用硫酸或各种氧化剂处理过的表面进行检测,结果表明硫酸在聚苯乙烯上产生的磺酸基团数量极少。该技术与各种使表面适用于细胞培养的氧化技术一起,产生了高表面密度的羟基。当这些基团被封闭时,黏附受到抑制,这证明了表面羟基对幼仓鼠肾细胞或白细胞黏附的重要性:羧基的封闭并未抑制黏附,反而可能提高表面的黏附力。这些结果适用于有血清和无血清时的细胞黏附。当用环己酰亚胺抑制蛋白质合成、用胰蛋白酶消化去除纤连蛋白并用亮抑蛋白酶肽终止胰蛋白酶活性后,细胞仍能在富含羟基的表面铺展,由此得出纤连蛋白对幼仓鼠肾细胞黏附并铺展到富含羟基表面的相对不重要性。