Q. Is additional L-glutamine or other supplementation required?
A. IS Sf Insect is a complete, ready-to-use medium. Additional supplementation is not required. However, it can be further supplemented according to user needs.
Q. What is the L-glutamine concentration in IS Sf Insect?
A. The medium contains 8 mM L-Glutamine.
Q. What is the glucose level in IS Sf Insect?
A. IS Sf Insect contains 8 gm/L glucose.
Q. Does the medium contain a sodium bicarbonate or other buffering system (if needed)?
A. There is no sodium bicarbonate in the medium since insect cells are typically cultured in 0% CO2. IS Sf Insect uses sodium phosphates as the buffering system.
Q. What are the recommendations for successful adaptation?
A. To assure optimum performance and allow for complete adaptation, maintain cells in IS Sf Insect for several passages, and passage twice a week at seeding density of 1.0 x 106 cells/mL to 2.0 x 106 cells/mL. A minimum of three passages in IS Sf Insect is strongly recommended before use in applications. Please refer to the product insert for recommended protocol.
Q. What do you advise to check for complete adaptation?
A. We use viability as the measurement and consider viability >90% as an indication of complete adaptation. Typically, the cells would reach that viability after three passages of adaptation. We suggest to passage the cells at higher density, for example 1.5 x 106 cells/mL, during the adaptation.
Q. What are the recommendations for adaptation if the cells are frozen in another medium?
A. To ensure successful adaption, thaw cells in the same medium which cells were banked in, and passage until viability >90% is achieved, then transfer the cells to IS Sf Insect. A minimum of three passages in IS Sf Insect medium is strongly recommended before use in applications.
Q. Is IS Sf Insect chemically defined?
A. IS Sf Insect is a serum-free and animal component-free medium. By our definition, it is not chemically-defined due to the low concentration of hydrolysates, or yeast extract, which may contain components that are not fully defined.
Q. What insect cell types does IS Sf Insect support?
A. IS Sf Insect is designed to support long-term, high-density suspension cultures of Sf9 and Sf21 cells.
Q. Can I freeze cells in IS Sf Insect?
A. Yes, we recommend cryopreservation of cells in 95% cold IS Sf Insect with 5% DMSO. Please refer to the product insert for protocol.