I treat PBMCs for 3 weeks with breast cancer conditioned media trying to differentiate them into active osteoclasts. Indeed, I produced TRAP positive multinucleated cells, but when I performed the same experiment on ivory slices they seem unable to resorb.
I do have a positive control treated with RANKL and the osteoclasts are able to resorb. Is there any chance to generate foreign body giant cells following treatment with cancer cells conditioned media?
Sometimes, TRAP positive cells haven't resorption capacity. The absent of integrins could be one of the reasons. Please, could you explain how did you prepare the medium? I am interested. Thank you!!!!
Thanks for your response Marcelo. I set up 4 million cancer cells in a T75 flask and the next day I replace their media with serum free fresh media. I collect the conditioned media after 24h and treat my PBMCs with 10% of it and 90% fresh media. Do you know whether these TRAP +ve cells without resorptive capacity are osteoclasts or it can be something else?
If they are taking 21 days to differentiate, you should allow them to resorb the surface for few more days. Certainly osteoclasts obtained after RANKL stimulation would be higher in number, more potent with distinct ruffled border, protease expression and acid secretion. Additionally you could try osteoassay plates. N.B. Not all breast cancer cells express RANKL. They might induce OSC formation through other cytokines such as TNF-a, IL-1.
This work can help ( like cells human osteoclasts MCP - 1-induced are tartrate -resistant acid phosphatase, NFATc1 , and receptor positive calcitonin but require activator receptor ligand NFkB bone resorption ) the fusion of precursors not It involves osteoclastogenesi
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology