Production of tissue plasminogen activator (t-PA) with differentiated F9-embryonic carcinoma cells grown as multicell spheroids

Abstract

F9 cells can only temporarily be cultivated as single cell suspension. The growth of multicell spheroids from a single cell suspension is shown. Within 24 hours the cells aggregated to multicell spheroids in spinner flasks. The aggregation was almost finished within 24 h and was independent of the initial cell concentration. but could be influenced by the geometry of the cultu re flask and the stirring velocity. A significant cell proliferation of the anchorage-dependent F9 cells was only detectable in the aggregated state. A narrow size distribution of multicell spheroids in a 3 days old culture revealed identical spheroid sizes. Under our conditions. with an inital cell density of 10 5 cells/ml a cell concentration of 6x10 6 cells/ml was reached within 4 days of spheroid culture. Cells in monolayers and in multicell spheroids were well coupled by gap junctions. Differentiated F9 cells showed a fibroblastoid morphology in contrast to the epitheloid morphology of undifferentiated F9 stem cells. This was not the case for multicell spheroids where the cells were also well coupled.