I'm planning to run energy metabolism assays (ATP, NAD/NADH) using HepG2 cells. The available cells are at passage 24. I’m aware that metabolic activity can change with higher passage numbers, potentially impacting assay reliability. Would it be advisable to consider a lower passage (e.g.,
Hello Shayal Devi,
For more accurate and reliable results from energy metabolism assays, I would suggest you use HepG2 cells within the first 20 passages. This will help to maintain the cell line's characteristics closer to its original phenotype.
Whenever cells are passaged, they undergo cellular drift and this drift can affect the cells' response to stimuli, making it difficult to interpret results in cell-based assays. Changes in their properties and characteristics such as alterations in morphology, gene expression, and metabolic activity are observed at higher passages. Whenever I have to perform cell-based assays, I use cells within passage 15, and I suggest you to follow the same.
For HepG2 cells it is generally recommended to maintain cells within a specific passage range, often between 8 and 20, for the most reliable performance in cell-based assays.
Best,
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