Standard 4-hours Chromium-51 (51Cr) Release Assay

Abstract

[Abstract] 51 Cr-release assays are commonly used for the precise and accurate quantification of cytotoxicity, particularly in the study of tumor cytolysis. This test has the advantage of requiring only very few cells. Materials and Reagents 1. Target cells: Acute Myeloid Leukemia (AML) cell line (U937 or HL60 from ATCC) or AML cells isolated from patients with AML 2. Effector cells: γδ T cells isolated from Healthy volunteer provided by Etablissement 3. Francais du Sang (EFS) or patients 4. Medium : RPMI 1640 (Life Technologies, Gibco ®) 5. Fetal calf serum (FCS) heat-inactivated 1 h at 56 °C 6. Chromium-51 (51 Cr, 5 mCi/ml) (PerkinElmer) 7. Complete medium = RPMI 1640 medium supplemented with 10% heat-inactivated FCS Equipment 1. Centrifuge for microplates 2. Microplates (96 well round-bottom) for cell incubations 3. LumaPlate TM (PerkinElmer) 4. Liquid scintillation counter 5. Incubator Controls 1. Spontaneous Release: Target cells were incubated alone (replace effector cells by 50 μl of media). After 4 h of incubation, wells were centrifuged and 50 μl of supernatant were recovered. 2. Maximum load: Target cells were incubated alone (replace effector cells by 50 μl of media). After 4 h of incubation, wells were mixed and 50 μl of cell suspension were