Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients

Autor: Lina Marcela Barrera, León Darío Ortiz, Hugo Grisales, Mauricio Rojas, Mauricio Camargo
Jazyk: English<br />Spanish; Castilian
Rok vydání: 2018
Předmět:
Zdroj: Biomédica: revista del Instituto Nacional de Salud, Vol 38, Iss 3, Pp 379-387 (2018)
Druh dokumentu: article
ISSN: 0120-4157
DOI: 10.7705/biomedica.v38i4.3882
Popis: Introduction: The quantification of chromosomal instability is an important parameter to assess genotoxicity and radiosensitivity. Most conventional techniques require cell cultures or laborious microscopic analyses of chromosomes or nuclei. However, a flow cytometry that selects the reticulocytes has been developed as an alternative for in vivo studies, which expedites the analytical procedures and increases up to 20 times the number of target cells to be analyzed. Objectives: To standardize the flow cytometry parameters for selecting and quantifying the micronucleated reticulocytesCD71+ (MN-RET) from freshly drawn peripheral blood and to quantify the frequency of this abnormal cell subpopulation as a measure of cytogenetic instability in populations of healthy volunteers (n =25), and patients (n=25), recently diagnosed with high-grade gliomas before the onset of treatment. Materials and methods: Blood cells were methanol-fixed and labeled with anti-CD-71-PE for reticulocytes, antiCD-61-FITC for platelet exclusion, and propidium iodide for DNA detection in reticulocytes. The MN-RETCD71+ cell fraction was selected and quantified with an automatic flow cytometer. Results: The standardization of cytometry parameters was described in detail, emphasizing the selection and quantification of the MN-RETCD71+ cellular fraction. The micronuclei basal level was established in healthy controls. In patients, a 5.2-fold increase before the onset of treatment was observed (p
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