Home >
Community >
How can I differentiate the THP-1 cells into osteoclasts?
Upvote
27
Downvote
+ Thp-1
+ Protocols
Posted by
Matt Robinson
How can I differentiate the THP-1 cells into osteoclasts?
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
Hi Ce Dou , would you mind telling me more details about the THP-1 differentiation protocol, like cell density and media used for osteoclasts differentiation? From my experience in RAW cells and other cells, cell density is important. Or anyone here have other suggestions? Thank you.
Hi Ce Dou , would you mind telling me more details about the THP-1 differentiation protocol, like cell density and media used for osteoclasts differentiation? From my experience in RAW cells and other cells, cell density is important. Or anyone here have other suggestions? Thank you.
Jenny, did you find any good protocol? I want to do the same differentiation in the (near) future and it would be very helpful to start from some protocol
Jenny, did you find any good protocol? I want to do the same differentiation in the (near) future and it would be very helpful to start from some protocol
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
Macrophages also express TRAP. While osteoclasts secrete both 5a and 5b, macrophages can't secret the 5b isoform, but its found intracellularly. There are different ways to differentiate between macrophages and osteoclasts, for example using a phagocytosis assay, staining for cathepsin K or assessing resorption.
Macrophages also express TRAP. While osteoclasts secrete both 5a and 5b, macrophages can't secret the 5b isoform, but its found intracellularly. There are different ways to differentiate between macrophages and osteoclasts, for example using a phagocytosis assay, staining for cathepsin K or assessing resorption.
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
More
VOTE
Hi Ce Dou , would you mind telling me more details about the THP-1 differentiation protocol, like cell density and media used for osteoclasts differentiation? From my experience in RAW cells and other cells, cell density is important. Or anyone here have other suggestions?
Thank you.
Hi Ce Dou , would you mind telling me more details about the THP-1 differentiation protocol, like cell density and media used for osteoclasts differentiation? From my experience in RAW cells and other cells, cell density is important. Or anyone here have other suggestions?
Thank you.
More
VOTE
Jenny, did you find any good protocol? I want to do the same differentiation in the (near) future and it would be very helpful to start from some protocol
Jenny, did you find any good protocol? I want to do the same differentiation in the (near) future and it would be very helpful to start from some protocol
More
VOTE
Hi, Ce Dou, would you mind tell me the difference between macrophage and osteoclast if THP-1 was used to differentiate.
Hi, Ce Dou, would you mind tell me the difference between macrophage and osteoclast if THP-1 was used to differentiate.
More
VOTE
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
Jenny and Irene, I personally have not done that before. But one of my colleague have successfully did that. Here is his protocol. THP-1 cells were cultured in DMEM media with 10% FBS, cells were first induced with PMA (50ng/mL) for 3-4 day for adhenrence. RANKL (50 ng/mL) and M-CSF (30 ng/mL) were then introduced for another 5-6 days. TRAP positive multinuleated osteoclast-like cells will occur. Concentration of PMA is the key to success induction. Hope this will do some help.
More
VOTE
Macrophages also express TRAP. While osteoclasts secrete both 5a and 5b, macrophages can't secret the 5b isoform, but its found intracellularly. There are different ways to differentiate between macrophages and osteoclasts, for example using a phagocytosis assay, staining for cathepsin K or assessing resorption.
Macrophages also express TRAP. While osteoclasts secrete both 5a and 5b, macrophages can't secret the 5b isoform, but its found intracellularly. There are different ways to differentiate between macrophages and osteoclasts, for example using a phagocytosis assay, staining for cathepsin K or assessing resorption.
More
VOTE
Osteoclasts should express TRAP and exhibit an actin ring.
A similar protocol is described here: https://www-ncbi-nlm-nih-gov.ezproxy.library.ubc.ca/pubmed/?term=PMID%3A+28983613
Osteoclasts should express TRAP and exhibit an actin ring.
A similar protocol is described here: https://www-ncbi-nlm-nih-gov.ezproxy.library.ubc.ca/pubmed/?term=PMID%3A+28983613
More
VOTE