Assessment of different protocols for the isolation and purification of gut associated lymphoid cells from the gilthead seabream (Sparus aurata L.).

Biological Procedures Online
Irene SalinasMaria Angeles Esteban

Abstract

Teleost gut associated lymphoid tissue (GALT) consists of leucocyte populations located both intraepithelially and in the lamina propria with no structural organization. The present study aims to assess different protocols for the isolation of GALT cells from an important fish species in the Mediterranean aquaculture, the gilthead seabream. Mechanical, chemical and enzymatic treatments were assayed. Nylon wool columns and continuous density gradients were used for further separation of cell subpopulations. Light microscopy and flow cytometry showed that the highest density band (HD) consisted of a homogeneous lymphocytic population, whereas the intermediate density band (ID) corresponded to epithelial and secretory cells and some lymphocytes. Respiratory burst activity of total cell suspensions revealed very low numbers of potential phagocytic cells, reflecting results from light microscopy and reports in other teleost species. The present data set up the basis for future functional characterization of GALT in seabream.

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Citations

Oct 3, 2012·Fish & Shellfish Immunology·Eve MartinJean-Pol Frippiat
Dec 3, 2011·Developmental and Comparative Immunology·Irene SalinasJ Oriol Sunyer
Oct 25, 2017·The Journal of Immunology : Official Journal of the American Association of Immunologists·Ali SepahiIrene Salinas
Nov 13, 2020·Frontiers in Cell and Developmental Biology·Youngjin ParkViswanath Kiron

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Methods Mentioned

BETA
light
flow cytometry
PMA
chemical treatment
dissection
scanning electron microscopy
flow
light microscopy

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