Percoll density gradient separation of cells from human malignant effusions

Abstract

Summary A simple method is described for the separation of cells derived from effusions of patients with adenocarcinomas in discontinuous density gradients of Percoll. After separation, cells from different fractions were analyzed by morphologic, histochemical and immunologic criteria. Total cell recovery from 27 experiments was 67+4%. Macrophages (82%) were recovered in the intermediate density fraction (1.056-1.067gml-1) with a purity of 90%. Recovered lymphocytes (98%) were found in the high density fraction (1.067-1.077gml-1) with a purity of 92%. The majority of the lymphocytes recovered were T cells. Malignant adenocarcinoma cells (90%) were recovered in the lowest density fractions (up to 1.056gml-1) with a purity of 79%. Use of effective cell separation procedures should facilitate the analysis of the functional capacities of both normal and neoplastic cells derived from human malignant effusions.

Tumor Cells in Blood Shed From the Surgical Field

“…Most of the negative results occurred with shed blood in which tumor cell numbers were below 104. In five of 16 samples in which invasiveness was tested in a Boyden chamber, tumor cells were found to be capable of actively passing the collagen type Ior type IV-coated Nucleopore mem¬ brane. Again, most negative results were observed with cell numbers below 104, in which the test system rarely iMicroscopic residual tumor (R1) was also present.…”

Differential T‐cell responses to a chimeric Plasmodium falciparum antigen; UB05‐09, correlates with acquired immunity to malaria

“…PBMCs for use in ELISpot assay were purified using percoll gradient method as earlier described . Briefly 8 mL of venous blood was collected from subjects and transferred into tubes containing EDTA anticoagulant.…”

Isolation of malignant cells from human bone marrow using a discontinuous percoll gradient

“…Band 3 preparations contain hematologic stem cells that might have the capacity to effect hematological reconstitution of a marrow-ablated patient [9,10]. This technique has been used in seven patients with malignant disease at this institution to prepare marrow harvested in remission for autologous bone marrow rescue (unpublished data).…”

“…Percoll gradients have also been shown to provide greater CFU-C enrichment in harvested bone marrow than techniques using Ficoll-paque [4]. Density gradients have been used to remove malignant cells from infiltrated organs [5-71 and effusions [8,9] in both animals and humans.…”