engleski

Study of different growth promotors on BHK 21 C13 biomass production

Summary: BHK 21 C13 cells were used as a model in production of vaccine against Aujeszky disease. In order to obtain sufficiant cell concentrations four different sera and growth promoter CABRPP were examined. CABRPP gave the best result and its application can significantly reduce the amount of necessary serum. Material and Methods: Baby hamster kidney cells (BHK 21 C13), ATCC CCl 10 stored at liquid N2 were used for biomass production. Cell propagation has been performed after thawing at 37 C with medium + 10 percent serum. Futher biomass production has been performed at T-flasks and cell spin bioreactor under controlled atmosphere. Influence of two different media (MEM-N and Glasgow BHK 21 medium), four different sera (horse serum, fetal calf serum, donor calf serum and newborn calf serum), three different initial cell concentrations and influence of growth promoter CABRPP were examined. Overview and discussion of results: During cell growth the total cell density, viability and growth kinetics were determined. The best result is obtained with Glasgow BHK 21 medium supplemented with 5 percent horse serum and initial cell concentration of 3.6x 10exp5. Total cell density was 1.6x 10exp6, viability over 95 percent and tD=16 h. In the case when the part of serum was replaced with growth promoter CABRPP while the total protein content remained the same total cell density increased for 50 percent (2.4x 10exp6), viability remained over 95 percent and tD reduced for 33 percent (tD 12 h). Vaccine production with such cells has shown the same or better activity when compared with traditional procedure. Conclusions: Addition of CABRPP increases total cell density (50 percent) and reduce tD (33 percent). Also CABRPP reduces the amount of necessary serum addition. References: (1) P.J.Radlett et al, Applied Microbiology, Oct.(1971). (2) P.Perin et al, Vaccine, vol.13 (13), (1995). (3) Kniewald et al, Vaccinology, today and tomorrow, Proceedings, (1995).

animal cells; high cell density; growth promoters; BHK 21 C13 cells

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