Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology

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The micronuclei test in lymphocyte culture is a validated procedure to study mutagenicity. It consists in detecting damaged interphasic nuclear material produced by chromosome fragmentation or nuclear division errors in 2000 binucleated cells with well defined cytoplasm. Standard protocol derives fr...

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Detalles Bibliográficos
Autores: Castillo, Erika, Guevara-Fujita, María Luisa, Fujita, Ricardo
Formato: artículo
Fecha de Publicación:2011
Institución:Universidad Nacional Mayor de San Marcos
Repositorio:Revistas - Universidad Nacional Mayor de San Marcos
Lenguaje:español
OAI Identifier:oai:ojs.csi.unmsm:article/241
Enlace del recurso:https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/241
Nivel de acceso:acceso abierto
Materia:biomonitoreo humano
Proyecto HUMN
genotoxicidad
citogenética
protocolo.
Human biomonitoring
HUMN Project
genotoxic
Cytogenetic
Protocol.
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spelling Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodologyOptimización del test de micronúcleos en linfocitos cultivados usando una metodología de gradiente y frotisCastillo, ErikaGuevara-Fujita, María LuisaFujita, Ricardobiomonitoreo humanoProyecto HUMNgenotoxicidadcitogenéticaprotocolo.Human biomonitoringHUMN ProjectgenotoxicCytogeneticProtocol.The micronuclei test in lymphocyte culture is a validated procedure to study mutagenicity. It consists in detecting damaged interphasic nuclear material produced by chromosome fragmentation or nuclear division errors in 2000 binucleated cells with well defined cytoplasm. Standard protocol derives from blood chromosome preparation with hypotonic and fixing solutions as well as preparing slides by dropping fixed cells. We modified the protocol to improve the number and quality of micronuclei. First, lymphocytes are separated from red cells before culture. After 72 hours, hypotonic and repeated fixer washing steps are eliminated. Cells are put onto slides by spreading instead of dropping. With these modifications we obtained more (about 8 times per slide) and better defined binucleated cells to help micronuclei detection.El test de micronúcleos en cultivo de linfocitos es una prueba validada para estudiar mutagenicidad. Consiste en detectar material nuclear interfásico dañado, producto de fragmentación cromosómica o errores de división nuclear en 2000 células binucleadas con citoplasma definido. El protocolo estándar deriva de la preparación citológica de cromosomas de sangre periférica con solución hipotónica, lavados de fijador y goteo de láminas. Nosotros proponemos modificaciones para mejorar el número y la observación de los micronúcleos. Primero se purifica linfocitos de glóbulos rojos antes del cultivo, luego de 72 horas, se elimina la solución hipotónica y los lavados repetidos con fijador, y finalmente se coloca las muestras en láminas por frotis en vez de goteo. Con las modificaciones obtenemos mas células binucleadas bien definidas (un promedio de 8 veces más por lámina) mejorando la eficiencia de este test.Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Biológicas2011-08-15info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/24110.15381/rpb.v18i2.241Revista Peruana de Biología; Vol. 18 Núm. 2 (2011); 261 - 263Revista Peruana de Biología; Vol. 18 No. 2 (2011); 261 - 2631727-99331561-0837reponame:Revistas - Universidad Nacional Mayor de San Marcosinstname:Universidad Nacional Mayor de San Marcosinstacron:UNMSMspahttps://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/241/228Derechos de autor 2011 Erika Castillo, María Luisa Guevara-Fujita, Ricardo Fujitahttps://creativecommons.org/licenses/by-nc-sa/4.0info:eu-repo/semantics/openAccessoai:ojs.csi.unmsm:article/2412020-05-21T13:00:35Z
dc.title.none.fl_str_mv Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
Optimización del test de micronúcleos en linfocitos cultivados usando una metodología de gradiente y frotis
title Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
spellingShingle Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
Castillo, Erika
biomonitoreo humano
Proyecto HUMN
genotoxicidad
citogenética
protocolo.
Human biomonitoring
HUMN Project
genotoxic
Cytogenetic
Protocol.
title_short Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
title_full Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
title_fullStr Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
title_full_unstemmed Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
title_sort Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
dc.creator.none.fl_str_mv Castillo, Erika
Guevara-Fujita, María Luisa
Fujita, Ricardo
author Castillo, Erika
author_facet Castillo, Erika
Guevara-Fujita, María Luisa
Fujita, Ricardo
author_role author
author2 Guevara-Fujita, María Luisa
Fujita, Ricardo
author2_role author
author
dc.subject.none.fl_str_mv biomonitoreo humano
Proyecto HUMN
genotoxicidad
citogenética
protocolo.
Human biomonitoring
HUMN Project
genotoxic
Cytogenetic
Protocol.
topic biomonitoreo humano
Proyecto HUMN
genotoxicidad
citogenética
protocolo.
Human biomonitoring
HUMN Project
genotoxic
Cytogenetic
Protocol.
description The micronuclei test in lymphocyte culture is a validated procedure to study mutagenicity. It consists in detecting damaged interphasic nuclear material produced by chromosome fragmentation or nuclear division errors in 2000 binucleated cells with well defined cytoplasm. Standard protocol derives from blood chromosome preparation with hypotonic and fixing solutions as well as preparing slides by dropping fixed cells. We modified the protocol to improve the number and quality of micronuclei. First, lymphocytes are separated from red cells before culture. After 72 hours, hypotonic and repeated fixer washing steps are eliminated. Cells are put onto slides by spreading instead of dropping. With these modifications we obtained more (about 8 times per slide) and better defined binucleated cells to help micronuclei detection.
publishDate 2011
dc.date.none.fl_str_mv 2011-08-15
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/241
10.15381/rpb.v18i2.241
url https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/241
identifier_str_mv 10.15381/rpb.v18i2.241
dc.language.none.fl_str_mv spa
language spa
dc.relation.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/rpb/article/view/241/228
dc.rights.none.fl_str_mv Derechos de autor 2011 Erika Castillo, María Luisa Guevara-Fujita, Ricardo Fujita
https://creativecommons.org/licenses/by-nc-sa/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Derechos de autor 2011 Erika Castillo, María Luisa Guevara-Fujita, Ricardo Fujita
https://creativecommons.org/licenses/by-nc-sa/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Biológicas
publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Ciencias Biológicas
dc.source.none.fl_str_mv Revista Peruana de Biología; Vol. 18 Núm. 2 (2011); 261 - 263
Revista Peruana de Biología; Vol. 18 No. 2 (2011); 261 - 263
1727-9933
1561-0837
reponame:Revistas - Universidad Nacional Mayor de San Marcos
instname:Universidad Nacional Mayor de San Marcos
instacron:UNMSM
instname_str Universidad Nacional Mayor de San Marcos
instacron_str UNMSM
institution UNMSM
reponame_str Revistas - Universidad Nacional Mayor de San Marcos
collection Revistas - Universidad Nacional Mayor de San Marcos
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1795238305132969984
score 13.754011
Improving the micronuclei test in cultured lymphocytes by gradient and cell spreading methodology
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