I would like to analyze cytokines secreted by arthritic PBMCs (human) after challenging them with collagen.

Is there a particular cell concentration and collagen concentration you would recommend?

- From papers I saw that 5million cells per mL cell concentration is used with 10-100ug/mL collagen. Seems to be typically in 24 or 48-well plates.

- However somebody that regularly does this with other antigens suggested that PBMCs are typically cultured at 1million cells per mL - this would greatly reduce our cell needs. I'm however a bit worried if the signal will be strong enough as collagen is supposedly a weak antigen.

- We would like to do this in 96-well format and 6 day incubation. We would analyze the supernatants for select cytokines at the end of incubation by Luminex.

Thank you all in advance for any recommendations!

More Ondrej Halgas's questions See All
Similar questions and discussions