Microculture Assay for Human Macrophage Maturation in vitro

Abstract

The terminal differentiation of monocytes (mo) to macrophages (MO) is essential for the complex functioning of the mononuclear phagocyte system. Similar to in vivo MO maturation, blood mo transform to MO when cultured in vitro. This offers a model system for experimental studies of functional defects within this cell lineage. In order to develop a microassay for the evaluation of MO maturation in vitro we monitored the antigenic phenotype of MO during differentiation in vitro. The cells were grown either in suspension on hydrophobic Teflon foils, or as plastic adherent monolayers. Surface antigens were visualized on single cells with the immunoperoxidase slide technique or quantitated with the enzyme-linked immunosorbent assay (ELISA). As MO mature in vitro they increase in cell size, in the amount of beta-2-microglobulin, the myeloid antigen CD14 and HLA-DR antigen expression as measured with cell-ELISA, both in microwell cultures and on single cells. Upon terminal differentiation mo-MO express differentiation-specific antigens (transferrin receptor, surface transferrin and antigens of the MAX series) which are indicative of a successful MO maturation. Monocyte cultures in microplates and the subsequent evaluation of differentiation antigen expression could be used as an experimental system to study the modulation of MO differentiation in vitro and may serve as an in vitro parameter of MO function in vivo.