In vitro antibody synthesis in 20 microliters hanging drops: initiation of secondary responses and a simple method of cloning hybridomas.
An in vitro culture system is described in which antibody synthesis is initiated by 10(5) hapten-primed spleen cells in 20 microliters hanging drops. Plaque-forming cell (PFC) responses to thymus dependent and thymus independent antigens were obtained in drop cultures comparable to the micro Mishell--Dutton system. Antigens could be administered in soluble form or on antigen-pulsed peritoneal cells. The hanging drop method also provides a reliable yet simple procedure for cloning hybridoma cells while permitting direct visualization of the number of cells in each drop.
D000818:Animals; D000917:Antibody Formation; D000955:Antigens, T-Independent / Q000276:immunology; D002478:Cells, Cultured; D003470:Culture Media; D006825:Hybridomas / Q000276:immunology*; D007156:Immunologic Memory; D007158:Immunologic Techniques; D051379:Mice
D000955:Antigens, T-Independent; D003470:Culture Media