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!