"Panning" for lymphocytes: a method for cell selection.

TLDR

The authors developed a simple method to fractionate T and B lymphocytes. The method uses antibody‑coated plastic dishes to selectively bind splenic B lymphocytes, allowing gentle removal of adherent cells while preserving function, and can detect and separate cell types as low as 2 % of the population or be adapted for double‑antibody selection. In experiments, 98 % of nonadherent cells were Ig negative and 97 % of adherent cells were Ig positive, demonstrating the method’s effectiveness for fractionating cells by surface antigens.

Abstract

We have developed a simple method for the fractionation of T and B lymphocytes. Plastic dishes coated with antibodies specific for mouse Ig selectively bind splenic B lymphocytes. The adherent cells are easily removed by gentle pipetting; both adherent and nonadherent populations retain immunologic function. In a typical experiment, when 3 X 10(7) splenic lymphocytes were added to a 100 X 15 mm plastic dish coated with microgram quantities of anti-Ig, 98 % of the nonadherent cells were Ig negative and 97% of the adherent cells were Ig positive. The method is sufficiently sensitive to allow detection and separation of cell types comprising as little as 2% of the total population and can be modified to allow the selection of cells by a double-antibody procedure. We believe that the plastic dish method will be generally useful for fractionating cells on the basis of their cell surface antigens.