title: Quantitative expansion of ES cell-derived myeloid progenitors capable of differentiating into macrophages.
abstract: Macrophages participate in physiologic and pathologic processes through elaboration of distinct activation programs. Studies with macrophage cell systems have revealed much concerning the importance of this pleiotropic cell; however, these studies are inherently limited by three factors: heterogeneity of the target cell population, poor capacity to elaborate various activation programs, and lack of a genetically tractable model system for loss- and gain-of-function studies. Although definitive, hematopoietic lineages can be isolated from embryonic stem (ES) cells, these isolation procedures are inefficient and time-consuming and require elaborate cell-sorting protocols. We therefore examined whether myeloid precursors, capable of differentiating into macrophages, could be conditionally expanded in vitro. Here, we report methods for selective isolation and immortalization of ES cell-derived myeloid precursors by estrogen-regulated HoxA9 protein. Using this new macrophage differentiation system, an unlimited number of custom-designed macrophages with defined functional characteristics can be generated from any targeted ES cell. In combination with knockout or small interfering RNA knockdown technologies, this macrophage differentiation system provides a powerful tool for high throughput analysis of regulatory mechanisms controlling macrophage activation in health and disease.
authors.affiliation: Division of Endocrinology, Metabolism and Gerontology, Department of Medicine and Graduate Program in Immunology, Stanford University School of Medicine, Stanford, CA 94305-5103, USA.
authors.forename: Justin I
authors.initials: JI
authors.lastname: Odegaard
authors.forename: Divya
authors.initials: D
authors.lastname: Vats
authors.forename: Lina
authors.initials: L
authors.lastname: Zhang
authors.forename: Roberto
authors.initials: R
authors.lastname: Ricardo-Gonzalez
authors.forename: Kristi L
authors.initials: KL
authors.lastname: Smith
authors.forename: David B
authors.initials: DB
authors.lastname: Sykes
authors.forename: Mark P
authors.initials: MP
authors.lastname: Kamps
authors.forename: Ajay
authors.initials: A
authors.lastname: Chawla
mesh_terms: D002454:Cell Differentiation / Q000276:immunology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000276:immunology*; D018398:Homeodomain Proteins / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology; D008262:Macrophage Activation / Q000276:immunology; D008264:Macrophages / Q000166:cytology / Q000276:immunology*; D023461:Myeloid Progenitor Cells / Q000166:cytology / Q000276:immunology*; D011960:Receptors, Estrogen / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology
chemical_list: D016207:Cytokines; D018398:Homeodomain Proteins; D011960:Receptors, Estrogen; C097899:homeobox protein HOXA9; D007371:Interferon-gamma