A simple method is described for preparing monolayers of non-adherent cells, using concanavalian A to bind the cells to wells of plastic microtest plates. The method was used successfully with all 202 human cell types tested, which included 23 tissue culture lines, 165 fresh specimens of all major histological types of leukemia and lymphoma, 20 fresh myelomas, 2 fresh thymomas, normal spleen and lymph node cells, fractionated T lymphocytes and B lymphocytes from peripheral blood, and cultured fetal amniotic cells. All cell types attached firmly, and were not detached by subsequent vigorous washing. In contrast, attempted attachment of cells in serum free medium, or with poly-L-lysine or glutaraldehyde, was ineffective with many cell types. We used the monolayers as target cells for antibodies to cell surface antigens, utilizing immune rosetting or complement-mediated cytotoxicity. This procedure should simplify most assays involving non-adherent target cells.
Bibliographical noteFunding Information:
I am grateful to Dr. Anthony Albino for the conjugated erythrocytes, to Ms. Catherine Higgins for technical assistance, to Drs. Kenneth O. Lloyd and Lloyd J. Old for advice and support, and to Yvette Johnson for preparing the manuscript. This work was supported in part by U.S. Public Health Service Grants CA-26184, CA 29989, CA-22507 and CA-08748, and a grant from the Borden Fund.
- cell surface antigens
- concanavalin A-mediated cell monolayer
- immune rosetting