Powdered M199 Medium

M199 is especially suitable for the culture of non-transformed cells and is also commonly used in virology, vaccine production, and in vitro culture of primary explants of rat pancreatic epithelial cells and mouse lens tissue. Compared with other basal media, M199 contains unique components, including adenine, adenosine, hypoxanthine, thymine, and other vitamins. M199 has two balanced salt compositions, Earle’s salt composition is commonly used in CO2 environment, Hank’s salt composition is used in non-CO2 environment.199 medium does not contain protein, lipid or growth factors. Medium 199 uses a sodium bicarbonate buffer system (2.2 g/L) and therefore requires a 5–10% CO2 environment to maintain physiological pH. For a wide range of cell culture applications, we provide modified M199 media with various components, as well as M199 media with customizable components.

Ingredients of Powdered M199 Medium

This M199 dry powder cell culture medium needs to be supplemented with sodium bicarbonate, adjusted for pH and filtered during preparation. The detailed preparation scheme is as follows:

1. Add 950 mL of distilled water to a mixing vessel as close to final volume as possible.
2. Add dry medium to room temperature (15°C to 30°C) water and stir gently. Do not heat water.
3. Rinse the inside of the pack to remove any residual powder.
4. Add 29.3 mL of 75 g/L NaHCO3 solution to the medium.
5. Adjust the pH to 0.2 to 0.3 units below the desired final working pH by slowly adding and stirring 1 N NaOH or 1 N HCl. The pH may rise by 0.1 to 0.3 units after filtration. The recommended working pH is 7.1-7.4.
6. Adjust final volume with distilled water.
7. Immediately process the medium through the membrane filtration function of a 0.2 μm filter into a sterile container using a positive pressure system.