Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients
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 h...
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doaj-5838ed5750fa4d58870aed2d269a45a42020-11-25T01:39:10ZengInstituto Nacional de SaludBiomédica: revista del Instituto Nacional de Salud0120-41570120-41572018-09-0138337938710.7705/biomedica.v38i4.38822145Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patientsLina Marcela Barrera0León Darío Ortiz1Hugo Grisales2Mauricio Rojas3Mauricio Camargo4Grupo de Genética, Regeneración y Cáncer (GRC), Instituto de Biología, Universidad de Antioquia, Medellín, ColombiaGrupo de Genética, Regeneración y Cáncer (GRC), Instituto de Biología, Universidad de Antioquia, Medellín, Colombia Instituto de Cancerología-Clínica Las Américas, Medellín, ColombiaGrupo de Investigación Demografía y Salud, Facultad Nacional de Salud Pública, Universidad de Antioquia, Medellín, ColombiaUnidad de Citometría de Flujo y Grupo de Inmunología Celular e Inmunogenética (GICIG), Facultad de Medicina, Universidad de Antioquia, Medellín, ColombiaGrupo de Genética, Regeneración y Cáncer (GRC), Instituto de Biología, Universidad de Antioquia, Medellín, ColombiaIntroduction: 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 <0.05). Conclusion: The data showed the usefulness of flow cytometry coupled with anti-CD-71-PE and anti-CD-61-FITC labeling in circulating reticulocytes as an efficient and high resolution method to quantify chromosome instability in vivo. Finally, possible reasons for the higher average of micronuclei in RETCD71+ cells from untreated high-grade glioma patients were discussed.https://www.revistabiomedica.org/index.php/biomedica/article/view/3882citometría de flujoreticulocitosinestabilidad cromosómicagliomapruebas de micronúcleos |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lina Marcela Barrera León Darío Ortiz Hugo Grisales Mauricio Rojas Mauricio Camargo |
spellingShingle |
Lina Marcela Barrera León Darío Ortiz Hugo Grisales Mauricio Rojas Mauricio Camargo Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients Biomédica: revista del Instituto Nacional de Salud citometría de flujo reticulocitos inestabilidad cromosómica glioma pruebas de micronúcleos |
author_facet |
Lina Marcela Barrera León Darío Ortiz Hugo Grisales Mauricio Rojas Mauricio Camargo |
author_sort |
Lina Marcela Barrera |
title |
Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients |
title_short |
Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients |
title_full |
Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients |
title_fullStr |
Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients |
title_full_unstemmed |
Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients |
title_sort |
flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients |
publisher |
Instituto Nacional de Salud |
series |
Biomédica: revista del Instituto Nacional de Salud |
issn |
0120-4157 0120-4157 |
publishDate |
2018-09-01 |
description |
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 <0.05).
Conclusion: The data showed the usefulness of flow cytometry coupled with anti-CD-71-PE and anti-CD-61-FITC labeling in circulating reticulocytes as an efficient and high resolution method to quantify chromosome instability in vivo. Finally, possible reasons for the higher average of micronuclei in RETCD71+ cells from untreated high-grade glioma patients were discussed. |
topic |
citometría de flujo reticulocitos inestabilidad cromosómica glioma pruebas de micronúcleos |
url |
https://www.revistabiomedica.org/index.php/biomedica/article/view/3882 |
work_keys_str_mv |
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