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

Salinas I, Meseguer J, Esteban MA - Biol Proced Online (2007)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Fish Innate Immune System Group, Department of Cell Biology, Faculty of Biology, University of Murcia, 30100 Murcia, Spain.

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.

No MeSH data available.


Related in: MedlinePlus

Fig. 2Scanning electron micrographs of nylon wool fibers from nylon wool columns incubated with seabream gut cells. (a) Mucus (arrow) from seabream gut trapped between nylon wool fibres. Bar: 200 μm (b) Small rounded cell (arrow head) adhered to a nylon wool fibre. Bar: 20 μm.
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f2: Fig. 2Scanning electron micrographs of nylon wool fibers from nylon wool columns incubated with seabream gut cells. (a) Mucus (arrow) from seabream gut trapped between nylon wool fibres. Bar: 200 μm (b) Small rounded cell (arrow head) adhered to a nylon wool fibre. Bar: 20 μm.

Mentions: Observation under scanning electron microscopy of nylon wool fibres from columns incubated for one hour with seabream gut cells, revealed considerable amounts of mucus attached to them (Fig. 2A). Additionally, some rounded small cells (diameter around 5μm) adhered to the fibres (Fig. 2B).


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

Salinas I, Meseguer J, Esteban MA - Biol Proced Online (2007)

Fig. 2Scanning electron micrographs of nylon wool fibers from nylon wool columns incubated with seabream gut cells. (a) Mucus (arrow) from seabream gut trapped between nylon wool fibres. Bar: 200 μm (b) Small rounded cell (arrow head) adhered to a nylon wool fibre. Bar: 20 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211574&req=5

f2: Fig. 2Scanning electron micrographs of nylon wool fibers from nylon wool columns incubated with seabream gut cells. (a) Mucus (arrow) from seabream gut trapped between nylon wool fibres. Bar: 200 μm (b) Small rounded cell (arrow head) adhered to a nylon wool fibre. Bar: 20 μm.
Mentions: Observation under scanning electron microscopy of nylon wool fibres from columns incubated for one hour with seabream gut cells, revealed considerable amounts of mucus attached to them (Fig. 2A). Additionally, some rounded small cells (diameter around 5μm) adhered to the fibres (Fig. 2B).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Fish Innate Immune System Group, Department of Cell Biology, Faculty of Biology, University of Murcia, 30100 Murcia, Spain.

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.

No MeSH data available.


Related in: MedlinePlus