A Guest Blog by Randall Alfano, Ph.D., Cell Culture Scientist, InVitria
It is well known that serum and other animal derived components can create inconsistency in cell culture, leading to variability in growth and performance. In order to reduce the amount of serum required, as well as the variability and final cost per liter of media, InVitria has developed Zap-SR. Zap-SR (Serum Reducer) is an animal free 100x cell culture supplement that is used to reduce the amount of serum required by up to 90% for many cell types, including hybridoma.
This study shows growth and performance data for hybridoma cells grown in 10% serum, reduced serum, and reduced serum + Zap-SR conditions. Significant decreases in growth and performance are seen in the reduced serum cultures where Zap-SR was not added, but it is clear that Zap-SR supplementation was able to recover the cells in both cases.
Two hybridoma cell lines were selected for this study. Hybridoma 1 preferred DMEM/F12 + GlutaMax while Hybridoma 2 was cultured in IMDM + GlutaMAX. Cells were expanded in the respective base media in addition to F-68 and 10% FBS in 50 mL culture volumes in 250 mL shake flasks.
To initiate the Zap-SR study, cells were seeded at an initial density of .45e5 cells/mL in 50 mL media containing either 10% serum, reduced serum, or reduced serum supplemented with 1x Zap-SR. Cells were cultured for 8 days in shake flask cultures at 37⁰C/5% CO2. Cultures were sampled every other day. Hybridomas cultured in reduced serum exhibited a reduced growth potential as evidenced by the day 4 peak cell densities of Hybridoma 1 and 2 (Figure A and B, respectively). The addition of 1x Zap-SR efficiently rescued hybridoma cell growth in both hybridoma lines.
To determine if equivalent cell growth yielded comparable hybridoma productivity, the supernatants of hybridoma 1 were analyzed for IgG levels via ELISA (Figure C). Zap-SR efficiently rescued the ability of Hybridoma 1 to that of 10% serum-supplemented cultures.
The cost effectiveness of Zap-SR is another key advantage of this supplement. Cost analysis comparing 10% serum-containing medium to reduced serum media supplemented with 1x Zap-SR demonstrates a 38 and 41% cost reduction for Hybridoma 1 and 2, respectively (Figure D).
Zap-SR is able to significantly reduce the amount of serum required to culture many cell lines, including hybridoma, providing many advantages for cell culture scientists.
InVitria develops, manufactures, and markets a portfolio of animal component free bioreagents and works to optimize media for a variety of different cell lines. These products are used in cell culture, biopharmaceutical formulation, Cell Therapy, regenerative medicine, medical devices and diagnostics. InVitria is the bioreagents division of Ventria Bioscience Inc. Learn more about InVitria’s products at www.InVitria.com.