Abstract

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 (1.056-1.067 g ml-1) with a purity of 90%. Recovered lymphocytes (98%) were found in the high density fraction (1.067-1.077 g ml-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 1.056 g ml-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.

Highlights

  • We have recently demonstrated autologous macrophages and lymphocytes modulate the growth of human tumour clonogenic cells (Hamburger & White, 1982; Hamburger et al, 1983a)

  • Fluids were centrifuged at 600g for 10min and cell pellets resuspended in McCoy's 5A medium containing 10% foetal bovine serum (FBS)

  • Twenty-four specimens used in this study were derived from patients with adenocarcinoma of the ovary (12), colon (5), breast (5), or unknown primary (2)

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Summary

Objectives

The aim of this study was to devise a method for separating these cells on density gradients of

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