title: New method to disaggregate and analyze single isolated helminthes cells using flow cytometry: proof of concept.
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.
authors.affiliation: Departamento de Infectología e Inmunología Perinatal, Instituto Nacional de Perinatología, 11000 México DF, Mexico.
authors.forename: Karen
authors.initials: K
authors.lastname: Nava-Castro
authors.forename: Romel
authors.initials: R
authors.lastname: Hernández-Bello
authors.forename: Saé
authors.initials: S
authors.lastname: Muñiz-Hernández
authors.forename: Galileo
authors.initials: G
authors.lastname: Escobedo
authors.forename: Jorge
authors.initials: J
authors.lastname: Morales-Montor
mesh_terms: D000818:Animals; D051242:Caveolin 1 / Q000032:analysis / Q000737:chemistry; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D015801:Helminth Proteins / Q000032:analysis / Q000737:chemistry; D051379:Mice; D010274:Parasitology / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D013552:Swine; D013621:Taenia / Q000737:chemistry / Q000166:cytology / Q000302:isolation & purification*; D013622:Taeniasis / Q000469:parasitology; D014234:Trichinella / Q000737:chemistry / Q000166:cytology / Q000302:isolation & purification*; D014235:Trichinellosis / Q000469:parasitology; D014335:Tropomyosin / Q000032:analysis / Q000737:chemistry
chemical_list: D051242:Caveolin 1; D015801:Helminth Proteins; D014335:Tropomyosin