Identification of progenitor cells in long-term spleen stromal cultures that produce immature dendritic cells

Dendritic cells (DC) are produced continuously by a unique, longterm culture (LTC) system in which hemopoiesis is supported by a splenic stromal cell layer in the absence of added growth factors. Flow cytometric analysis reveals the production of two distinct cell subsets. The more predominant large-cell subset resembles highly endocytic DC that are large, granular, and possess membrane extensions. They also express high levels of the DC markers CD11c, CD11b, DEC-205, and CD80 on their cell surface. They do not resemble mature DC because they express low levels of MHC type II and CD86 molecules, as well as c-kit and Fc receptor (FcR). These are known characteristics of immature DC. Small cells are smaller and less granular than large cells, with negative to low expression of CD11c, DEC-205, and CD86. A majority of small cells express varying levels of CD11b and CD80. Subpopulations of small cells express low levels of c-kit, FcR, and MHC type II, and only a 20% subpopulation is weakly endocytic. Upon transfer to an irradiated stromal layer, cells within the small subset proliferate and differentiate to resemble the large cells in size, complexity, membrane extensions, and CD11c and CD86 expression. The two cell subsets produced in LTC are developmentally linked, with the heterogeneous small-cell subset containing progenitors of the larger homogeneous, immature DC subset. LTC represent a valuable model system for studying DC development from hemopoietic progenitors.