iv) Wild-type vs XID mice in 16 h ZIP model
Male CBA/CaCrl (WT) or XID mice (8-10 weeks old) were given zymosan
challenge (100 ug; i.p.). Blood was collected by direct cardiac
puncture, peritoneal exudate cells were harvested in ice-cold PEC
harvest buffer (PBS, 5 mM EDTA) and spleen dissected 16 h after zymosan
challenge.
Murine bone marrow‐derived
macrophages
(BMDMs):
Bone marrow‐derived macrophages were generated as previously described
(Recio et al., 2018). Briefly, fresh bone marrow cells from tibiae and
femurs of male C57BL/6 or XID mice aged 8–10 weeks were cultured in
DMEM containing 4.5 g·L−1 glucose, 2 mM
l‐glutamine, 50 units·ml−1 penicillin and 50
μg·ml−1 streptomycin, 10% heat‐inactivated FBS, 10%
L929 cell‐conditioned media (as a source of macrophage
colony‐stimulating factor) and for 7 days. Bone marrow cells were seeded
into 8 ml of medium in 90 mm non‐tissue culture treated Petri dishes
(ThermoFisher Scientific, Sterilin, UK). On Day 5, an additional 5 ml of
medium was added. Gentle scraping was used to lift cells. BMDMs were
then counted and suspended in FBS free media at the desired cell
concentration.