Fractionation of human tumour-associated cells by centrifugal elutriation to establish lymphocyte, macrophage and tumor cell cultures from a single tumour biopsy.
The application of the Beckman centrifugal elutriator system for the simultaneous isolation of lymphocytes, macrophages and tumour cells from enzyme disaggregated tumour-associated cells or from peritoneal aspirates is described. Using this technique we have been able to prepare cultures of these cells from single human tumour biopsies. Overall cellular recovery was always in excess of 70% and the purity of cell fractions varied between 89 and 99%. Lymphocyte fractions were found to be free of tumour cell contamination and this allowed lymphocyte cultures in T-cell growth factor containing medium to be established. Using this approach it should prove possible to investigate the clonal effector functions of tumour infiltrating lymphocytes against autologous and allogeneic tumour cells.
D000230:Adenocarcinoma / Q000473:pathology; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002498:Centrifugation; D003470:Culture Media; D005260:Female; D006801:Humans; D007376:Interleukin-2; D007958:Leukocyte Count; D008214:Lymphocytes / Q000473:pathology; D008264:Macrophages / Q000473:pathology; D009369:Neoplasms / Q000473:pathology*; D009504:Neutrophils / Q000473:pathology; D010051:Ovarian Neoplasms / Q000473:pathology
D003470:Culture Media; D007376:Interleukin-2