Nature Methods 3, 287 - 293 (2006)
Published online: 22 March 2006; | doi:10.1038/nmeth865
Quantitative production of macrophages or neutrophils ex vivo using conditional Hoxb8Gang G Wang1, 2, Katherine R Calvo1, 3, Martina P Pasillas1, David B Sykes1, 4, Hans Häcker5
& Mark P Kamps11
Department of Pathology & Molecular Pathology Graduate Program, School of Medicine, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA. 2
Biomedical Sciences Graduate Program, School of Medicine, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA. 3
Laboratory of Pathology, National Health Institutes, National Cancer Institute, 10 Center Drive, Bldg. 10, 2N212, Bethesda, Maryland 20892, USA. 4
Department of Medicine, Harvard Massachusetts General Hospital, 55 Fruit Street, Boston Massachusetts 02114, USA. 5
St. Jude Children's Research Hospital, Department of Infectious Diseases, Room E8062, 332 North Lauderdale Street, Memphis, Tennessee 38105, USA.
Correspondence should be addressed to Mark P Kamps mkamps@ucsd.edu Differentiation mechanisms and inflammatory functions of neutrophils and macrophages are usually studied by genetic and biochemical approaches that require costly breeding and time-consuming purification to obtain phagocytes for functional analysis. Because Hox oncoproteins enforce self-renewal of factor-dependent myeloid progenitors, we queried whether estrogen-regulated Hoxb8 (ER-Hoxb8) could immortalize macrophage or neutrophil progenitors that would execute normal differentiation and normal innate immune function upon ER-Hoxb8 inactivation. Here we describe methods to derive unlimited quantities of mouse macrophages or neutrophils by immortalizing their respective progenitors with ER-Hoxb8 using different cytokines to target expansion of different committed progenitors. ER-Hoxb8 neutrophils and macrophages are functionally superior to those produced by many other ex vivo differentiation models, have strong inflammatory responses and can be derived easily from embryonic day 13 (e13) fetal liver of mice exhibiting embryonic-lethal phenotypes. Using knockout or small interfering RNA (siRNA) technologies, this ER-Hoxb8 phagocyte maturation system represents a rapid analytical tool for studying macrophage and neutrophil biology.
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