Author: Cyktor, Joshua C.; Carruthers, Bridget; Beamer, Gillian L.; Turner, Joanne
Title: Clonal Expansions of CD8(+) T Cells with IL-10 Secreting Capacity Occur during Chronic Mycobacterium tuberculosis Infection Document date: 2013_3_5
ID: 1rhlu59c_12
Snippet: Mice were euthanized by CO 2 asphyxiation and lungs perfused with cold phosphate buffered saline containing 50 Units/mL of heparin through the right ventricle of the heart. Lungs from individual mice were mechanically disrupted using a GentleMACS dissociator (Miltenyi Biotec, Boston, MA) followed by collagenase A (type XI) (0.7 mg/mL, Sigma) and type IV bovine pancreatic DNAse (30 mg/mL, Sigma) digestion at 37uC for 30 minutes in GentleMACS C-tub.....
Document: Mice were euthanized by CO 2 asphyxiation and lungs perfused with cold phosphate buffered saline containing 50 Units/mL of heparin through the right ventricle of the heart. Lungs from individual mice were mechanically disrupted using a GentleMACS dissociator (Miltenyi Biotec, Boston, MA) followed by collagenase A (type XI) (0.7 mg/mL, Sigma) and type IV bovine pancreatic DNAse (30 mg/mL, Sigma) digestion at 37uC for 30 minutes in GentleMACS C-tubes. Lung cell suspensions were passed through a 70 mm nylon cell screen and residual erythrocytes were lysed with Gey's solution. Viable cells were determined by trypan blue exclusion.
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