In vitro production of monoclonal antibodies to cultured embryonic chick limb mesenchyme.

Abstract

A simple, rapid protocol for the in vitro production of monoclonal antibodies (MAbs) that recognize native antigens in cultured chick limb mesenchyme during chondrogenic differentiation is described. Murine lymphocytes were stimulated by direct exposure to methanol-fixed micromass cultures of limb mesenchyme derived from the distal tip of stage 25 chick limb buds. Initial immunohistochemical characterization of two antibodies (DIDI and DIIA5) produced by this method showed preferential localization of reactivity with antigens in developing cartilage nodules during chondrogenesis in cultured chick limb mesenchyme. This study demonstrates the utility of in vitro immunization of lymphocytes for the production of MAbs to native antigens expressed by differentiating embryonic limb cells in culture. Immunohistochemical data provided by DIDI and DIIA5 suggest that antigens bearing these epitopes may be important in early morphogenetic events during limb skeletal development.