Abstract
Supernatants or conditioned media (CM) were produced by rainbow trout pronephros cells (PNC) and spleen cell cultures (1×106/ml), by addition of 20 μg/ml phythaemagglutinin (PHA), 5 ng/ml 12-O-tetradecanoyl-phorbol-13-acetate (PHA) (for 2h), PHA together with PMA, 10% horse serum (HS), or 100 μg/ml concanavalin A (ConA) after a culture of 7 days. Only PNC were effective growth factor producers. The effect of different concentrations of CM (5%–28%) on cell number was tested after a cultivation period of 14 days. PNC at a concentration of 1×106/ml were cultured with various concentrations of CM and the proliferation was tested by the XTT-test (testing the dehydrogenase activity of the cells by formation of a formazan) after 10 days. CM produced by cells with PHA, PHA and PMA and HS increased the proliferation in a concentration-dependent manner. CM produced with PMA alone was effective in the XTT-test. There was no synergistic or additive effect of PMA with PHA. CM produced with ConA had no effect, although in the XTT-test a strong proliferation of PNC in presence of 100 μg/ml ConA was observed. In a semisolid culture with collagen, CM treatment resulted in prevention of cell death and an increase in cell size but did not induce proliferation. All CM obtained from spleen cells had no effect. Stimulation of spleen cells by CM could be seen only in the XTT-test. PHA and HS trigger the PNC to release growth factors in vitro which stimulate cell growth and/or prevent cell death.
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Siegl, E., Hlawa, A., Blunk, H. et al. Pronephros and spleen cells from rainbow trout (Oncorhynchus mykiss) in vitro: Growth factors produced under the influence of mitogens. Comparative Haematology International 5, 261–267 (1995). https://doi.org/10.1007/BF02044143
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DOI: https://doi.org/10.1007/BF02044143