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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ftdoajarticles:oai:doaj.org/article:5838ed5750fa4d58870aed2d269a45a4 2023-05-15T15:12:09+02:00 Flow cytometry in peripheral blood reticulocytes as a marker of chromosome instability in highgrade glioma patients Lina Marcela Barrera León Darío Ortiz Hugo Grisales Mauricio Rojas Mauricio Camargo 2018-09-01T00:00:00Z https://doi.org/10.7705/biomedica.v38i4.3882 https://doaj.org/article/5838ed5750fa4d58870aed2d269a45a4 EN ES eng spa Instituto Nacional de Salud https://www.revistabiomedica.org/index.php/biomedica/article/view/3882 https://doaj.org/toc/0120-4157 0120-4157 doi:10.7705/biomedica.v38i4.3882 https://doaj.org/article/5838ed5750fa4d58870aed2d269a45a4 Biomédica: revista del Instituto Nacional de Salud, Vol 38, Iss 3, Pp 379-387 (2018) citometría de flujo reticulocitos inestabilidad cromosómica glioma pruebas de micronúcleos Medicine R Arctic medicine. Tropical medicine RC955-962 article 2018 ftdoajarticles https://doi.org/10.7705/biomedica.v38i4.3882 2022-12-31T01:02:02Z 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. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Biomédica 38 3 379 387 |
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English Spanish |
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citometría de flujo reticulocitos inestabilidad cromosómica glioma pruebas de micronúcleos Medicine R Arctic medicine. Tropical medicine RC955-962 |
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citometría de flujo reticulocitos inestabilidad cromosómica glioma pruebas de micronúcleos Medicine R Arctic medicine. Tropical medicine RC955-962 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 |
topic_facet |
citometría de flujo reticulocitos inestabilidad cromosómica glioma pruebas de micronúcleos Medicine R Arctic medicine. Tropical medicine RC955-962 |
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. |
format |
Article in Journal/Newspaper |
author |
Lina Marcela Barrera León Darío Ortiz Hugo Grisales Mauricio Rojas Mauricio Camargo |
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 |
publishDate |
2018 |
url |
https://doi.org/10.7705/biomedica.v38i4.3882 https://doaj.org/article/5838ed5750fa4d58870aed2d269a45a4 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Biomédica: revista del Instituto Nacional de Salud, Vol 38, Iss 3, Pp 379-387 (2018) |
op_relation |
https://www.revistabiomedica.org/index.php/biomedica/article/view/3882 https://doaj.org/toc/0120-4157 0120-4157 doi:10.7705/biomedica.v38i4.3882 https://doaj.org/article/5838ed5750fa4d58870aed2d269a45a4 |
op_doi |
https://doi.org/10.7705/biomedica.v38i4.3882 |
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Biomédica |
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38 |
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379 |
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387 |
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