A new approach for rapid and reliable enumeration of circulating endothelial cells in patients
| Autor: | M. A. den Bakker, Jaco Kraan, Stefan Sleijfer, Jan-Willem Gratama, Cornelis Verhoef, Anieta M. Sieuwerts, Michiel Strijbos, John A. Foekens |
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| Přispěvatelé: | Medical Oncology, Pathology, Surgery |
| Rok vydání: | 2012 |
| Předmět: |
Pathology
medicine.medical_specialty Genotype CD34 Antigens CD34 Cell Count CD146 Antigen Biology Real-Time Polymerase Chain Reaction Flow cytometry Immunophenotyping Antigen Predictive Value of Tests Neoplasms Enumeration medicine Humans Netherlands medicine.diagnostic_test Reverse Transcriptase Polymerase Chain Reaction Endothelial Cells Reproducibility of Results Hematology Flow Cytometry Prognosis Molecular biology Immunohistochemistry Real-time polymerase chain reaction Phenotype Gene Expression Regulation Case-Control Studies cardiovascular system CD146 Leukocyte Common Antigens Biomarkers |
| Zdroj: | Journal of Thrombosis and Haemostasis, 10(5), 931-939. Wiley-Blackwell Publishing Ltd |
| ISSN: | 1538-7836 1538-7933 |
| Popis: | To cite this article: Kraan J, Strijbos MH, Sieuwerts AM, Foekens JA, den Bakker MA, Verhoef C, Sleijfer S, Gratama JW. A new approach for rapid and reliable enumeration of circulating endothelial cells in patients. J Thromb Haemost 2012; 10: 931–9. Summary. Background: Mature circulating endothelial cells (CECs) are surrogate markers of endothelial damage/dysfunction. A lack of standardized assays and consensus on CEC phenotype has resulted in a wide variation of reported CEC numbers (4–1300 per mL). Objectives: Given the need for a quick, reliable, robust and validated CEC assay at an affordable price, we present a novel approach to enumerate CECs using a multi-parameter flow cytometric (FCM) method without immunological pre-enrichment. Methods: CECs were defined as CD34+, CD45neg, CD146+ and DNA+ events based on the immunophenotype of endothelial cells from vein-wall dissections. As CECs express high levels of CD34, we based our assay on absolute CD34 counts after analyzing all CD34 positive events in a total blood volume of 4 mL needed for a precise enumeration of CECs at a frequency of < 1 cell lL )1 . Results: The endothelial origin of CECs was confirmed by morphology, immunohistochemistry and gene expression. The new FCM assay was tested in parallel with a validated assay (i.e. CellSearch � ). CEC levels ranged from 4 to 79 CEC mL )1 in healthy individuals and were significantly higher in patients with advanced solid malignancies (P = 0.0008) and in patients with hematological malignancies (P < 0.0001). Conclusions: This flow cytometric method should be useful as a fast and economical assay to enumerate and characterize CECs. |
| Databáze: | OpenAIRE |
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