Varani J, Dame M, Rediske J, Beals T F, Hillegas W
J Biol Stand. 1985 Jan;13(1):67-76. doi: 10.1016/s0092-1157(85)80035-4.
Normal diploid human fibroblasts and first passage monkey kidney epithelial cells were examined for growth and metabolic activity on microcarriers made from glass and on microcarriers made from DEAE-dextran. The cells grew to a higher density (cells cm2 of surface area) on the glass microcarriers made from glass and on microcarriers made from DEAE-dextran. The cells grew to a higher density (cells/cm2 of surface area) on the glass microcarriers than they did on the DEAE-dextran microcarriers and morphological differences were observed between the cells growing on the two substrates. On the DEAE-dextran microcarriers, the cells were much more resistant to protease-mediated detachment than were the cells on the glass microcarriers. In these respects, the cells grown on the glass microcarriers were similar to cells grown in conventional monolayer culture. Interestingly, the cells grown on the DEAE-dextran microcarriers expressed higher levels of proteolytic enzyme activity than the cells grown on the glass microcarriers. Substrate-dependent differences in prostaglandin production also occurred--both in unstimulated cells and in cells stimulated with 12-0-tetradecanoyl phorbol acetate. The unstimulated cells on the glass microcarriers produced slightly higher levels of three different prostaglandins than did the cells on the DEAE-dextran microcarriers. However, after stimulation the levels were much higher in the DEAE-dextran microcarrier cultures than in the glass microcarrier cultures. In contrast to these results, there was no significant, substrate-dependent difference in the production of infectious herpes simplex virus. Taken together, these findings suggest that when commercially-useful cells such as normal fibroblasts and epithelial cells are grown in large quantities on microcarriers, the nature of the substrate may have a profound effect on the growth and physiology of the cells. They also suggest that when microcarriers are used, unexpected results based on preliminary work in conventional monolayer culture may be obtained.
对正常二倍体人成纤维细胞和第一代猴肾上皮细胞在由玻璃制成的微载体以及由二乙氨基乙基葡聚糖（DEAE - 葡聚糖）制成的微载体上的生长和代谢活性进行了检测。细胞在由玻璃制成的微载体和由DEAE - 葡聚糖制成的微载体上生长至更高的密度（每平方厘米表面积的细胞数）。细胞在玻璃微载体上比在DEAE - 葡聚糖微载体上生长至更高的密度（每平方厘米表面积的细胞数），并且观察到在两种基质上生长的细胞之间存在形态差异。在DEAE - 葡聚糖微载体上，细胞比在玻璃微载体上的细胞对蛋白酶介导的脱离更具抗性。在这些方面，在玻璃微载体上生长的细胞类似于在传统单层培养中生长的细胞。有趣的是，在DEAE - 葡聚糖微载体上生长的细胞比在玻璃微载体上生长的细胞表达更高水平的蛋白水解酶活性。在前列腺素产生方面也出现了底物依赖性差异——在未刺激的细胞以及用12 - O - 十四酰佛波醇 - 13 - 乙酸酯刺激的细胞中均如此。在玻璃微载体上的未刺激细胞产生的三种不同前列腺素水平略高于在DEAE - 葡聚糖微载体上的细胞。然而，刺激后，DEAE - 葡聚糖微载体培养物中的水平比玻璃微载体培养物中的水平高得多。与这些结果相反，在感染性单纯疱疹病毒的产生方面不存在显著的底物依赖性差异。综上所述，这些发现表明，当诸如正常成纤维细胞和上皮细胞等具有商业用途的细胞在微载体上大量生长时，基质的性质可能对细胞的生长和生理学产生深远影响。它们还表明，当使用微载体时，可能会获得基于传统单层培养初步工作的意外结果。
