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Neutrophil Lifespan Extension with CLON-G and an In Vitro Spontaneous Death Assay

Article DOI: 10.3791/65132-v • 05:52 min • May 12th, 2023
May 12th, 2023 •
*1,2, *3, *4, 1,2, 5, 1,2, 5
1State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, 2Tianjin Institutes of Health Science, 3Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, 4Haihe Laboratory of Cell Ecosystem, Tianjin Medical University, 5Joint Program in Transfusion Medicine, Department of Pathology, Harvard Medical School; Division of Blood Bank, Department of Laboratory Medicine, The Stem Cell Program, Boston Children's Hospital, Dana-Farber /Harvard Cancer Center
* These authors contributed equally

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This protocol details the preparation of CLON-G to extend the neutrophil lifespan to greater than 5 days and provides a reliable procedure for evaluating neutrophil death with flow cytometry and confocal fluorescence microscopy.

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Neutrophil Lifespan Extension CLON-G In Vitro Spontaneous Death Assay Cocktail Medium Neutrophil Functions Dialysis Preservation Transportation Gene Manipulation RPMI 1640 FBS Pen-strep HSP 70 G-CSF Centrifuge Tube Neutrophil Culture Non Tissue Cultured Plates Cell Suspension Incubate Stain Cytospin Light Microscope Flow Cytometry Analysis
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