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

Authors

  • Erika Castillo 1 Centro de Genética y Biología Molecular. Facultad de Medicina Humana, Universidad de San Martín de Porres, Alameda del Corregidor 1531, La Molina, Lima, Perú. Tel. 511 3652300 anexo 152. 2 Facultad de Ciencias Biológicas, Universidad Nacional Mayor de San Marcos
  • María Luisa Guevara-Fujita Centro de Genética y Biología Molecular. Facultad de Medicina Humana, Universidad de San Martín de Porres, Alameda del Corregidor 1531, La Molina, Lima, Perú. Tel. 511 3652300 anexo 152.
  • Ricardo Fujita Centro de Genética y Biología Molecular. Facultad de Medicina Humana, Universidad de San Martín de Porres, Alameda del Corregidor 1531, La Molina, Lima, Perú. Tel. 511 3652300 anexo 152.

DOI:

https://doi.org/10.15381/rpb.v18i2.241

Keywords:

Human biomonitoring, HUMN Project, genotoxic, Cytogenetic, Protocol.

Abstract

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.

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Published

08/15/2011

Issue

Vol. 18 No. 2 (2011)

Section

Notes

License

Copyright (c) 2011 Erika Castillo, María Luisa Guevara-Fujita, Ricardo Fujita

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How to Cite

Castillo, Erika, María Luisa Guevara-Fujita, and Ricardo Fujita. 2011. “Improving the Micronuclei Test in Cultured Lymphocytes by Gradient and Cell Spreading Methodology”. Revista Peruana De Biología 18 (2): 261-63. https://doi.org/10.15381/rpb.v18i2.241.