Varani J, Dame M, Beals T F, Wass J A
Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Biotechnol Bioeng. 1983 May;25(5):1359-72. doi: 10.1002/bit.260250515.
Three established cell lines were examined for growth on a newly developed microcarrier which consists of glass beads. The cells were simultaneously examined for growth on commercially available microcarriers made from DEAE-dextran and from plastic. Cell yields on the glass microcarriers were comparable to the cell yields on the commercially available products. Cells grown on the glass microcarriers were easily separated from the substratum by trypsinization (as were the cells grown on the plastic substratum) while the cells grown on the DEAE-dextran particles were much more trypsin resistant. After removal of cells from the glass microcarriers, the cells reattached and spread out in plastic flasks as readily as cells harvested from monolayer. Scanning electron microscopy revealed dramatic differences in the appearance of the cell grown on the glass microcarriers and cells grown on the DEAE-dextran microcarriers. On the glass microcarriers, cells attached to the substratum through lond, slender filopodia while on the DEAE-dextran microcarriers, the entire edge of the cell appeared to be in contact with the substratum. This dissimilarity in attachment could underly the difference in sensitivity to trypsin-mediated detachment. Finally, the glass microcarriers were washed after being used once and retested for their ability to support cell growth a second time. Nearly identical results were obtained with the reprocessed beads as with previously unused ones.
对三种已建立的细胞系在一种新开发的由玻璃珠组成的微载体上的生长情况进行了检测。同时检测了这些细胞在由DEAE-葡聚糖和塑料制成的市售微载体上的生长情况。玻璃微载体上的细胞产量与市售产品上的细胞产量相当。在玻璃微载体上生长的细胞通过胰蛋白酶消化很容易与基质分离(在塑料基质上生长的细胞也是如此),而在DEAE-葡聚糖颗粒上生长的细胞对胰蛋白酶的抗性要强得多。从玻璃微载体上移除细胞后,这些细胞重新附着并像从单层收获的细胞一样容易在塑料培养瓶中铺展。扫描电子显微镜显示,在玻璃微载体上生长的细胞与在DEAE-葡聚糖微载体上生长的细胞外观有显著差异。在玻璃微载体上,细胞通过长而细的丝状伪足附着于基质,而在DEAE-葡聚糖微载体上,细胞的整个边缘似乎都与基质接触。这种附着方式的差异可能是对胰蛋白酶介导的脱离敏感性不同的原因。最后,玻璃微载体使用一次后进行清洗,并再次测试其支持细胞生长的能力。再处理后的珠子获得的结果与之前未使用过的珠子几乎相同。