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Blood, 1 February 2004, Vol. 103, No. 3, pp. 852-859. Prepublished online as a Blood First Edition Paper on October 2, 2003; DOI 10.1182/blood-2003-04-1030. This Article Full Text (PDF) All Versions of this Article: 2003-04-1030v1 103/3/852    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Lieber, J. G Articles by Worthen, G S. Search for Related Content PubMed PubMed Citation Articles by Lieber, J. G Articles by Worthen, G S. Social Bookmarking                   What's this? Submitted April 3, 2003 Accepted September 16, 2003 The in vitro production and characterization of neutrophils from embryonic stem cells Jonathan G Lieber*, Saiphone Webb, Benjamin T Suratt, Scott K Young, Gary L Johnson, Gordon M Keller, and G Scott Worthen Department of Medicine, National Jewish Medical & Research Center, Denver, CO, USA Department of Immunology, National Jewish Medical & Research Center, Denver, CO, USA Department of Medicine, University of Colorado, Denver, CO, USA Institute for Gene Therapy and Molecular Medicine, Mount Sinai School of Medicine, New York, NY, USA Department of Pharmacology, University of Colorado, Denver, CO, USA * Corresponding author; email: lieberjg{at}njc.org. An embryonic stem (ES) cell/OP9 coculture system for the effective production of functional neutrophils is described. A three step differentiation strategy was developed that utilizes liquid culture, enabling reliable and abundant production of neutrophils at high purity without the need of sorting for isolation of mature neutrophils. Utilization of the OP9 stromal cell line significantly enhances the number, percentage and duration of differentiated neutrophils produced from embryonic stem cells. Effective and sustained differentiation of ES cells into neutrophils provides a useful model system for studying neutrophil differentiation and function and the factors that regulate them. Morphological and functional evaluation of these ES-derived neutrophils indicates that large numbers of mature neutrophils can be produced from totipotent ES cells in vitro. Specifically, their morphology, ability to produce superoxides, flux calcium and chemotax in response to MIP-2, stain for the granulocyte specific marker specific chloroacetate esterase, and contain the neutrophil specific markers Gr-1 and the mouse neutrophil specific antigen indicates that they are comparable to purified mouse bone marrow neutrophils. They also express gelatinase and lactoferrin granule proteins. During the differentiation of these ES-derived neutrophils, regional areas of neutrophil production can be identified that have been designated as neutrophil generating regions (NGRs). CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: D. van der Hoeven, T. C. Wan, and J. A. Auchampach Activation of the A3 Adenosine Receptor Suppresses Superoxide Production and Chemotaxis of Mouse Bone Marrow Neutrophils Mol. Pharmacol., September 1, 2008; 74(3): 685 - 696. [Abstract] [Full Text] [PDF] P. 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