Fc block treatment, dead cells exclusion, and cell aggregates discrimination concur to prevent phenotypical artifacts in the analysis of subpopulations of tumor-infiltrating CD11b(+) myelomonocytic cells
| Autor: | Curzio Rüegg, François Kuonen, Cédric Touvrey, Julien Laurent |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Histology
Angiogenesis Cell Bone Marrow Cells Cell Separation Receptors Fc Biology Monocytes Pathology and Forensic Medicine Flow cytometry Mice Lymphocytes Tumor-Infiltrating Cell Line Tumor medicine Animals Receptor Cell Aggregation Mammary tumor Mice Inbred BALB C medicine.diagnostic_test Cell Death CD11 Antigens Mammary Neoplasms Experimental Cell Biology Flow Cytometry Molecular biology medicine.anatomical_structure Cancer cell Syngenic Female Artifacts Cytometry |
| Zdroj: | Cytometry. Part A : the journal of the International Society for Analytical Cytology. 77(11) |
| ISSN: | 1552-4930 |
| Popis: | It is well established that cancer cells can recruit CD11b(+) myeloid cells to promote tumor angiogenesis and tumor growth. Increasing interest has emerged on the identification of subpopulations of tumor-infiltrating CD11b(+) myeloid cells using flow cytometry techniques. In the literature, however, discrepancies exist on the phenotype of these cells (Coffelt et al., Am J Pathol 2010;176:1564-1576). Since flow cytometry analysis requires particular precautions for accurate sample preparation and trustable data acquisition, analysis, and interpretation, some discrepancies might be due to technical reasons rather than biological grounds. We used the syngenic orthotopic 4T1 mammary tumor model in immunocompetent BALB/c mice to analyze and compare the phenotype of CD11b(+) myeloid cells isolated from peripheral blood and from tumors, using six-color flow cytometry. We report here that the nonspecific antibody binding through Fc receptors, the presence of dead cells and cell doublets in tumor-derived samples concur to generate artifacts in the phenotype of tumor-infiltrating CD11b(+) subpopulations. We show that the heterogeneity of tumor-infiltrating CD11b(+) subpopulations analyzed without particular precautions was greatly reduced upon Fc block treatment, dead cells, and cell doublets exclusion. Phenotyping of tumor-infiltrating CD11b(+) cells was particularly sensitive to these parameters compared to circulating CD11b(+) cells. Taken together, our results identify Fe block treatment, dead cells, and cell doublets exclusion as simple but crucial steps for the proper analysis of tumor-infiltrating CD11b(+) cell populations. (C) 2010 International Society for Advancement of Cytometry |
| Databáze: | OpenAIRE |
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