Isolation and Culture of Human and Murine Neutrophils

Cultures of T84 IECs were grown as described previously (26 (link), 38 (link), 73 (link)). Human PMN were isolated from whole blood obtained from healthy male and female volunteers, with approval from the University of Michigan IRB on human subjects, using a previously described Polymorphprep density gradient centrifugation technique (73 (link), 74 (link)). Isolated PMN were 98% pure and > 95% viable and were used for all described assays within 2 hours of blood draw. Murine neutrophils were isolated from bone marrow extracted from the femur and tibias of male and female C57BL/6 mice using EasySep kits from Stem Cell Technologies as previously described (75 (link)).