A method to study apoptosis in eosinophils by flow cytometry
| Autor: | Per Venge, Lena Håkansson, Agneta Lukinius, Kristina Sandström |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Programmed cell death
Pathology medicine.medical_specialty Light Immunology Population Apoptosis DNA Fragmentation Phosphatidylserines Biology Flow cytometry Necrosis chemistry.chemical_compound Annexin medicine Humans Scattering Radiation Immunology and Allergy fas Receptor Propidium iodide Annexin A5 Sodium Azide education Cells Cultured education.field_of_study Staining and Labeling medicine.diagnostic_test Eosinophil Flow Cytometry Antigens Differentiation Molecular biology Staining Eosinophils medicine.anatomical_structure chemistry Propidium |
| Zdroj: | Journal of Immunological Methods. 240:55-68 |
| ISSN: | 0022-1759 |
| Popis: | The aim of this study was to develop a simple flow cytometric procedure to study eosinophil apoptosis. Eosinophils were isolated from the peripheral blood of healthy, non-allergic individuals and then cultured in basal culture medium. The cells were examined after 24, 48 and 72 h for forward- and side scatter (FS-SSC) pattern, staining with FDA, PI, and anti-CD95, and light microscopic appearance. After culture for >24 h, two populations with different FS-SSC-patterns appeared, referred to as A and B. Population A consisted of living, FDA-positive eosinophils. The eosinophils in population B showed a lower FS scatter than those in population A and a staining pattern with PI indicating the presence of hypodiploid DNA. Anti-CD95 demonstrated a significant staining of the eosinophils in population B, which increased after 2 days in culture. The cells were sorted using a FACS-Scan cell sorter and by Annexin V-coated magnetic beads to permit separate analyses of PI-staining pattern, DNA electrophoresis, and light microscopic examination of the cells in population B. The present study suggest that it is possible to discriminate between apoptotic and living eosinophils using the FS-SSC pattern and the PI-staining pattern obtained by flow cytometry. |
| Databáze: | OpenAIRE |
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