yes, its necessary to add in culture media for any type of cells. Because L-glutamine is needed for cell viability, proliferation and most required when cells are dividing rapidly.

Hello Matt Mannino

Yes, it would be necessary to add glutamine to McCoy's media. The explanation is provided below.

Both Caki-1 and Caki-2 are human clear cell renal cell carcinoma (ccRCC) cell lines. In ccRCC, there is a loss of VHL and fructose-1,6-bisphosphatase 1 (FBP1) tumor suppressor functions which results in an increased HIF activity. HIF reprograms cellular metabolism and increases glucose uptake and glycolysis via increased expression of glucose transporters and glycolytic enzymes.

Moreover, HIF also shifts the TCA cycle from predominant glucose utilization to a predominantly glutamine-fueled system. In normal cells, glucose-derived carbons enter the mitochondria via pyruvate, which drives the TCA cycle in a ‘clockwise’ fashion. However, even though ccRCC cells show increased glycolysis, increased HIF activity leads to inhibition of pyruvate dehydrogenase and pyruvate carboxylase activity, thus resulting in drastically decreased entry of glucose-derived carbon into the TCA cycle.

In consequence, these cells fuel the TCA cycle primarily in a counterclockwise direction by glutamine-derived glutamate via reductive carboxylation of aKG to isocitrate. Glutamine-addicted cancer cells operate the TCA cycle in this ‘reverse’ direction to generate citrate and malate for lipid biosynthesis, and oxaloacetate for nucleotide biosynthesis, while deriving most of their energy from glycolysis.

Therefore, HIF activation in ccRCC is both necessary and sufficient for increased glutamine utilization and dependence, both by limiting the ability of glucose to fuel the TCA cycle and by reprogramming glutamine metabolism to provide the requisite macromolecules needed for sustaining the increased needs of rapidly dividing cells.

You may want to refer to the below attached article for more information.

Article The ‘Achilles Heel’ of Metabolism in Renal Cell Carcinoma: G...

Best.

Hey Malcolm Nobre

Thank you! is there a concentration in particular you would suggest?

Thank you again

It is 2mM L-Glutamine.

Malcolm Nobre I found some McCoy's Modified 5a medium from ATCC that has 1.5 mM L-Glutamine. Would 1.5 mM be sufficient for growing Caki-1 and Caki-2 or should I manually supplement more into the media?

Thank you!

No need to supplement more. 1.5mM L-Glutamine should be sufficient.