Isolation and characterization of extracellular vesicles in Candida albicans

  1. Virginia Pérez-Doñate 1
  2. Facundo Pérez-Giménez 1
  3. Lucas del Castillo Agudo 1
  4. Juan Alberto Castillo-Garit 1
  5. Mar Soria-Merino 2
  6. Eulogio Valentín Gómez 1
  1. 1 Universitat de València
    info

    Universitat de València

    Valencia, España

    ROR https://ror.org/043nxc105

  2. 2 Hospital Clinico Universitario de Valencia
    info

    Hospital Clinico Universitario de Valencia

    Valencia, España

    ROR https://ror.org/00hpnj894

Revista:
Nereis: revista iberoamericana interdisciplinar de métodos, modelización y simulación

ISSN: 1888-8550

Año de publicación: 2020

Número: 12

Páginas: 99-108

Tipo: Artículo

DOI: 10.46583/NEREIS_2020.12.611 DIALNET GOOGLE SCHOLAR lock_openDialnet editor

Otras publicaciones en: Nereis: revista iberoamericana interdisciplinar de métodos, modelización y simulación

Resumen

Background: The occurrence of systemic infections due to C. albicans has increased especially in critically ill patients. In fungal infections, secretory mechanisms are key events for disease establishment. Recent findings demonstrate that fungal organisms release many molecular com-ponents to the extracellular space in extracellular vesicles. Aims: We develop a method to obtain exosomes from yeast cultures of the Candida albicans. Methods: Yeast strains used in this work were C. albicans SC5314, C. parapsilosis (ATCC 22019) and C. krusei (ATCC 6258). Yeasts were grown at 37.º in liquid YPD medium. The cell cultures were centrifuged and the supernatant filtered through sterile nitrocellulose. Filtrates were concentrated and centrifuged using an ultracentrifuge. The sediment was analyzed by electron microscopy of transmission.Results: The transmission of electron microscopy and nanoparticle tracking analysis confirmed the presence of extracellular vesicles (exosomes) of sizes between 100 and 200 nm and the absence of cellular contaminants. This was ratified by the characterization of proteins performed through the western blot technique, where the absence of cell contamination in the preparations was assessed. Conclusions: The method proves to be highly effective due to the homogeneity and purity of the obtained microvesicles. The protocol developed in this paper proves to be effective for obtaining exosomes of other Candida species, which will allow future studies to determine its protein composition and the role that these vesicles can play.

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Fuente de los datos: Dialnet