Dear Dr. Muhammad Hafeez

L-glutamine is a necessary supplement when culturing Sf9 cells in Grace's Insect Medium, especially for suspension cultures.

Sf9 cells, like other insect cells, rely on a complex nutrient medium for growth and protein production. L-glutamine serves as a primary source of nitrogen and carbon for cells and plays a critical role in various metabolic pathways, including the TCA cycle, which is essential for energy production.

The specific concentration of L-glutamine required may vary depending on the cell density, culture conditions and other factors. For culturing Sf9 cells, the typical L-glutamine concentration in culture medium is 2-4 mM. This is the most common range for L-glutamine concentration in Sf9 cell culture media. However, for high cell density cultures, you may have to use higher concentrations such as 6-8 mM to ensure sufficient glutamine is available.

You should use GlutaMAX compared to freshly prepared L-glutamine because unlike L-glutamine, GlutaMAX does not spontaneously degrade into ammonia or other toxic compounds during storage or in culture. GlutaMAX, due to its dipeptide structure prevents degradation into toxic byproducts like ammonia. This stability ensures consistent nutrient availability and avoids detrimental effects on cell health. There is a controlled release of L-glutamine from GlutaMAX ensuring a steady supply of this essential nutrient for cell growth and metabolism.

So, you should supplement Grace’s Insect Medium, which does not contain L-glutamine with GlutaMAX.

Regards,

Malcolm Nobre