A study on in situ high density cultivation of anchorage dependent animal cells.

在普通方法中，锚定依赖的动物细胞在固体培养基质上培养，如页岩或微珠的表面。这种处理可能有有害的影响，如细胞膜蛋白的蛋白水解。为了避免这些影响，本研究采用液-液界面系统进行细胞培养。细胞生长在培养基和疏水液体之间的界面上。在界面处培养的动物细胞的扩散比固体表面情况下要小，但该系统可以具有原位条件。在本研究中，得到以下结果：(1)表面活性剂的数量和种类对细胞在界面处的粘附有影响。(2)细胞形态受表面活性剂浓度控制，表面活性剂的浓度可改变细胞对界面的粘附。(3)细胞在界面处的扩散相对较小，使得界面处的细胞锚定。更有规律的半球形聚集体，形状像一个球体。(4)也尝试了在乳化液表面培养，结果发现乳化液的直径分布要均匀，这样才能使细胞恢复，防止彼此合并^<**>。(5)乳状液与油包水和油包水体系的培养效率均高于平板。^<**>Sato， M., Hishinuma， T., Terachi， K., Hatori， T.和Saito， M.：“蒸化水中绝缘和粘性液体的静电单分散”，静电，1995（第9届国际会议），P1.21, pp.65-66（纽约，1995）；Sato， M., Hatori, t .和Saito， M.：“液-液系统中静电分散液滴形成机制的实验研究”，Conf.Rec。ias物质。Meet (IEEE), pp.1485-1492（奥兰多，1995）；Sato， M., Kato, s.和Saito， M.：“使用交流会聚电场生产O/W型均匀尺寸液滴”，IEEE Trans。Ind.Appl。， 32 138-145 (1996)；Sato， M., Kudo， N.和Saito， M.，“应用振动射流方法的外加电场降低液体的表面张力”，Conf.Rec。ias Annu.Meet。（IEEE）， pp.1762-1767 (San Diego, October, 1996

In an ordinary method, anchorage-dependent animal cells cultivated on a solid culture substrate, such as the surface of shale or micro-beads. This treatment may have detrimental effects such as the proteolysis of the cell membrane proteins. To avoid these effects, cell cultivation using a liquid-liquid interface system was used in the present study. The cells grow at the interface between a culture medium and a hydrophobic liquid. The animal cells cultivating at the interface spread less than the case of solid surface but the system Could have in situ conditions. In the present study, the following results were obtained :(1) Quantity and kind of the surface active agent affected to the adhesion of the cells at the interface.(2) Morphology of the cell was controlled by concentration of surface active agent which varied cell adhesion to the interface.(3) Cell spreading at the interfaces was relatively small, so that the cells at an interface anchored. with one another and formed multicel … More lular hemispherical aggregations shaped like a spheroid.(4) Cultivation on the emulsion surface was also tried, as a result, it was found that the diameter distribution of the emulsion should be uniform for the purpose of recovery of the cells and preventing the coalescence of each other^<**>.(5) Using emulsion with the systems of oil-in water and water-in-oil, cultivation efficiency was exceeded than the case of using flat plate in both cases.^<**>Sato, M., Hishinuma, T., Terachi, K., Hatori, T and Saito, M. : "Electrostatic mono dispersion of insulating and viscous liquid in distillled water, "Electrostatics'95 (9th International Conference), P1.21, pp.65-66 (York, 1995) ; Sato, M., Hatori, T.and Saito, M. : "Experimental investigation of droplet formation mechanisms by electrostatic dispersion in a liquid-liquid system, "Conf.Rec.-IAS Annu.Meet (IEEE), pp.1485-1492 (Orlando, 1995) ; Sato, M., Kato, S.and Saito, M. : "Production of O/W type uniformly sized droplets using AC convergent eletric field, "IEEE Trans. Ind.Appl., 32,138-145 (1996) ; Sato, M., Kudo, N., and Saito, M., "Surface tension reduction of liquid by applied electric field using vibrating jet method, "Conf.Rec.-IAS Annu.Meet. (IEEE), pp.1762-1767 (San Diego, October, 1996) Less