GROWTH AND MORPHOLOGY OF ANCHORAGE-DEPENDENT ANIMAL-CELLS IN A LIQUIDLIQUID INTERFACE SYSTEM/

In general, anchorage-dependent animal cells cultivated on a solid cul ture substrate, such as polystyrene, are collected by trypsin treatmen t. This treatment may have detrimental effects such as the proteolysis of the cell membrane proteins. To avoid these effects, cell cultivati on using a liquid/liquid interface system has been investigated. In th is cultivation method, the cells grow at the interface between a cultu re medium and a hydrophobic liquid. In this study, various fluorocarbo ns (FC-40, FC-70, KPF-91, KPF-102, and KPF-142) were used as substrate s for the interface, and the cultivation of fibroblast cells (L-929; t he mouse-derived cell line) at the interfaces was investigated. Early in the cultivation period, the growth of L-929 cells depended on the s ubstrate type. Although cell cultivation at the interfaces was possibl e, it was slower than that at the polystyrene surface. Cell spreading at the interfaces was relatively small, which indicates that cell adhe sion at the interfaces may be weak. In particular, the cells at the ME M/FC-70 interface anchored with one another and formed multicellular h emispherical aggregations shaped like spheroids. The difference in the adhesions to the interfaces appears to be dependent on the contaminan ts contained in the fluorocarbons because the physical properties of t he fluorocarbon did not affect the cell growth and adhesion. Moreover, subcultivation from the interfaces to the same interface was possible without trypsin treatment. In this case, the delay of the growth at t he interfaces did not occur because the cells were not affected by try psin treatment. (C) 1998 John Wiley & Sons, Inc.