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Journal of Biomedicine and Biotechnology
Volume 2011 (2011), Article ID 257060, 9 pages
http://dx.doi.org/10.1155/2011/257060
Research Article

New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept

1Departamento de Infectología e Inmunología Perinatal, Instituto Nacional de Perinatología, 11000 México DF, Mexico
2Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP 70228, 04510 México DF, Mexico
3Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Secretaría de Salud (SSA), 47000 México DF, Mexico

Received 23 June 2011; Accepted 5 September 2011

Academic Editor: Luis I. Terrazas

Copyright © 2011 Karen Nava-Castro et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells) is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells.