In vitro stimulation of mouse spleen cells by mouse urine

Urinary tract-associated lymphoid structures (UTLASs), tertiary lymphoid tissues, are formed in renal pelvis (RP) of humans and mice with chronic kidney disease. We found that UTALS development was accelerated by urine leakage from RP lumen to the parenchyma following dysfunction of transitional epithelium covering UTLASs. Thus, UTALS-forming cells were stimulated by urine including urinary bio active substances. The spleens were isolated from MRL/MpJ at 6-month-old, minced, washed, resuspended, and cultured. These cultured cells were stimulated with urine supernatant derived from MRL/MpJ or MRL/MpJ-Faslpr/lpr mice or with PBS for 3 hours. And then, RNA was collected and gene expression was analyzed.