The in vitro production and characterization of neutrophils from embryonic stem cells
| Autor: | G. Scott Worthen, Benjamin T. Suratt, Gordon Keller, Saiphone Webb, Scott K. Young, Gary L. Johnson, Jonathan G. Lieber |
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| Rok vydání: | 2004 |
| Předmět: |
Stromal cell
Neutrophils Cellular differentiation Immunology MAP Kinase Kinase Kinase 1 Cell Separation Biology Biochemistry Cell Line Mice Superoxides Neutrophil differentiation medicine Animals Gelatinase Mice Knockout Stem Cells Cell Differentiation Chemotaxis Cell Biology Hematology MAP Kinase Kinase Kinases Molecular biology Embryonic stem cell Coculture Techniques Chemotaxis Leukocyte medicine.anatomical_structure Cell culture Mutation Tetradecanoylphorbol Acetate Bone marrow Stromal Cells |
| Zdroj: | Blood. 103:852-859 |
| ISSN: | 1528-0020 0006-4971 |
| DOI: | 10.1182/blood-2003-04-1030 |
| Popis: | An embryonic stem (ES) cell/OP9 coculture system for the effective production of functional neutrophils is described. A 3-step differentiation strategy was developed that uses liquid culture, enabling reliable and abundant production of neutrophils at high purity without the need of sorting for isolation of mature neutrophils. Use 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. Morphologic and functional evaluation of these ES-derived neutrophils indicates that large numbers of mature neutrophils can be produced from pluripotent ES cells in vitro. Specifically, their morphology, ability to produce superoxides, flux calcium, undergo chemotaxis in response to macrophage inflammatory protein 2 (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 with 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). |
| Databáze: | OpenAIRE |
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