After inducing THP- cells with PMA 200ng/ml for 24 hours, I took photos of the cells at 48 hours, 72 hours, 80 hours, and 96 hours. At 96 hours, the bright areas inside the cells seemed to have vacuoles. Does this mean that the cells are in a bad state?
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Lucian Miles,
Yes, the cells are in a bad state. PMA is a potent activator of protein kinase C, which is involved in various cellular processes, including differentiation. In THP-1 cells, PMA triggers a cascade of events that ultimately lead to their differentiation into macrophage-like cells.
PMA induces differentiation in a time-dependent manner. Short-term PMA treatment (24-48 hours) is commonly used for differentiation. However, prolonged exposure (beyond 80 - 96 hours) can cause excessive PKC activation, leading to cellular damage, including the formation of vacuoles.
So, whenever you use PMA for THP-1 differentiation, it is essential to monitor the cells closely and avoid prolonged exposure as it can negatively impact cell health and cause unwanted morphological changes.
Best,
If the cells induced by PMA have some morphological trends and the cells have attached to the wall, it means that PMA has a certain induction effect.
The vacuoles produced in the cells may be caused by the cell stress state. Generally, they look a bit like lipid droplets. And PMA treatment may affect the lipid metabolism of the cells, leading to the accumulation of lipid droplets. You can continue to culture and observe, detect the expression of surface CD11b, and judge the cell induction effect.
If the optimization differentiation results are not as expected, you need to review the initial state of the cells and adjust the concentration gradient of PMA.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
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