I wanted to know whether PMA treatment of Thp1 cells activates them or differentiates them. Certain literature suggest that PMA treatment for 24hours activate the cells following which it can be differentiated into the M1 and M2 subtypes using different stimuli. Can someone explain to me the difference between the "activation" and "differentiation"? When we say "activated", does it mean that the cells are committed/primed to become macrophages? Also, can different concentrations of PMA impact the extent of activation and subsequent differentiation?
PMA treatment of THP-1 monocytic cells activate protein kinase C signaling and leads to differentiation into macrophages ( enlargement of cells , elongation of cells , adherence of cells) by arresting cell proliferation. At this time, if you measure ERK it will not show differences between test and control. you have to rest them for 48 hours post differentiation to measure the reality of test product/or antigen. Activated THP-1 cells generally mean activation with IFN-gamma and LPS.
As replied above. Activation is non specific. Differentitaion requires cytokines.
To be clear, when the Thp1 cells are treated with PMA does it activate it or can that be considered to be a differentiated state?
it is both but not specific activation in terms of cell signaling responses due to PMA. what is that you are looking for?. if I know your experimental interest i can suggest whether this will have any impact.
I should have been more clear. PMA differentiates the monocytes into macrophages. Activation (to be stimulated) usually requires some thing, like LPS or any other stimulants. PMA treated macrophages do produce some cytokines, but you can push for more with stimulation. I usually don't consider PMA treated macrophages as "activated", but that has to do with the model I was working in. However, using a combination of IFNg and IL-4 and others, depending on the cocktail, you can shift the macrophages into a M1 or M2.
Dinesh Kumar Parandhaman I am interested in studying the process of monocyte to macrophage differentiation. I am not clear whether PMA induces the differentiation or do I have to use different cytokines to get them differentiated.
Shen-An Hwang Okay. So can I consider PMA treated Thp1s as differentiated macrophages?
Yes, you can. However, just understand that in vivo monocytes differentiate into macrophages only (I believe) when they migrate into tissue. At this point, they are kind of both being stimulated and differentiated at the same time.
So I'm not sure you can completely separated out differentiation without some sort of activation as everyone mentioned above.
Hi Dipsikha,
monocytes derived from blood can undergo natural differentiation. there is a procedure called panning where PBMCs are plated in culture plates/or dishes for 3 hours/or overnight followed by washing away. the adherent cells are called macrophages and they are allowed to mature for 5 days before any infection or stimulation is done.
- Badges
- Science method