Dear all,
Recently I plan to test a protein for its potential chemoattractant property for macrophages chemotaxis. And I plan to test on both mouse and human cell.
I realized some used RAW264.7 for mouse cell and THP-1 for human cell.
Can I use them too? Moreover, literally the THP-1 is the monocyte, can I use it for macrophage chemotaxis. If not, what other choices?
Ps. I am working on muscle and skeletal system.
Thank you so much.
Best Regards.
For mouse you may use RAW264.7 cell lines but why dont to try isolating BMDC from mouse femur. If you have access to animal facility and indent few mouse.
For Human isolating Monocytes is not difficult (we do it regularly). You should isolate monocytes from human peripheral blood and differentiate these into macrophage by stimulators (MCSF/PMA). You will get sufficient numbers of monocytes to be converted into macrophage, to be used in your chemotaxis assay.
Alternatively you may use THP-1 cell line (which is not a real alternative to monocytes as suggested by Prof. James). But you should differentiate the THP-1 into macrophage as well to carry out your research.
Thanks.
Hello,
You can stimulate ThP-1 with 50 ng of PMA for 48 hours to obtain macrophages. In this way, you Will be able to use them for macrophage chemotaxis.
There is the question of chemotaxis by leukemia cells versus macrophages. If you isolate monocytes from PB and culture them in M-CSF you can have a culture of macrophages to study. Note, THP-1 cells are used as a model of monocytes, but are not monocytes. Very easy to isolate monocytes and differentiate into macrophages. Regardless of what you study be aware of limitations.
Ikram Omar Osman Thank you so much Ikram. It really helps and I will try. Can I ask if you have any suggestions for macrophage cell line in mouse which is suitable for chemotaxis assay?
Thank you again.
James E Talmadge Thank you so much Dr. James. I will try it. And do you have any suggestions for macrophage cell line in mouse which is suitable for chemotaxis assay?
Thank you so much again.
@ziming Chen, I usually use BMDMs cells, when I do studies on murine macrophages. You can try to grow them, but I can't tell you if that would answer the question of your purpose.
Ikram Omar Osman Thank you Ikram so much for your patience and answer. I will try it.
Best regards.
I agree with the use of BMDM. Easy to isolate and differentiate. One femur will work well and provide a number of macs.
James E Talmadge Thank you so much Dr. James. I will try it.
Best regards.
Hi there, I think both options are fine. We previously used RAW264.7 cells. You can also use THP but differentiate with the use of PMA.
https://www.atcc.org/products/all/TIB-71.aspx
For mouse you may use RAW264.7 cell lines but why dont to try isolating BMDC from mouse femur. If you have access to animal facility and indent few mouse.
For Human isolating Monocytes is not difficult (we do it regularly). You should isolate monocytes from human peripheral blood and differentiate these into macrophage by stimulators (MCSF/PMA). You will get sufficient numbers of monocytes to be converted into macrophage, to be used in your chemotaxis assay.
Alternatively you may use THP-1 cell line (which is not a real alternative to monocytes as suggested by Prof. James). But you should differentiate the THP-1 into macrophage as well to carry out your research.
Thanks.
Jitendra Kumar Shandilya Thank you Jitendra so much for your detailed answer. I just start this project and the ethic document already took me too long time. So I want to test it on cell lines in advance. Your answer is very helpful. I will try it on primary cell in the future.
Thank you again.
Best regards.