Yamaji H, Fukuda H
Engineering Research Laboratories, Kaneka Corporation, Hyogo, Japan.
Appl Microbiol Biotechnol. 1992 May;37(2):244-51. doi: 10.1007/BF00178179.
Growth and death of anchorage-independent animal cells entrapped within porous biomass support particles (BSPs) in static or shake-flask cultures were evaluated by comparison of enzyme activity with non-immobilized cells grown under static culture using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and release of lactate dehydrogenase into the culture medium. Mouse myeloma MPC-11 (ATCC CCL 167) cells inoculated within porous polyvinyl formal resin BSPs (3 x 3 x 3 or 2 x 2 x 2 mm; mean pore diameter, 60 microns) grew exponentially at a specific growth rate comparable to that of non-immobilized cells in the initial period of incubation. Entrapped cells then reached the stationary phase with a cell density over 10(7) cells/cm3 BSP. The death rate of entrapped cells increased in response to the rise in viable cell density in the BSPs. Observation of viable cell distribution within the BSPs using MTT staining indicated that the cells concentrated within a thin outer shell of the BSPs with time. After the immobilized cells reached the stationary phase, penetration of cells into the outer shell ceased and heterogeneous distribution of cell density occurred in the viable cell layer in the shake-flask culture.
通过使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法比较酶活性以及将乳酸脱氢酶释放到培养基中的情况，对静态或摇瓶培养中包埋在多孔生物质支持颗粒（BSPs）内的非贴壁动物细胞的生长和死亡进行了评估。接种在多孔聚乙烯醇缩甲醛树脂BSPs（3×3×3或2×2×2毫米；平均孔径60微米）内的小鼠骨髓瘤MPC-11（ATCC CCL 167）细胞在培养初期以与非固定化细胞相当的比生长速率呈指数增长。然后，包埋的细胞达到稳定期，细胞密度超过10(7)个细胞/立方厘米BSP。包埋细胞的死亡率随着BSPs中活细胞密度的增加而上升。使用MTT染色观察BSPs内活细胞的分布表明，随着时间的推移，细胞集中在BSPs的薄外壳内。固定化细胞达到稳定期后，细胞向外壳的渗透停止，摇瓶培养中活细胞层出现细胞密度的不均匀分布。
