Optimization of the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss)

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F19%3AN0000158" target="_blank" >RIV/00027162:_____/19:N0000158 - isvavai.cz</a>

Result on the web

<a href="https://fvhe.vfu.cz/files/Sbornik_KMVP_2019.pdf" target="_blank" >https://fvhe.vfu.cz/files/Sbornik_KMVP_2019.pdf</a>

DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Optimization of the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss)

Original language description

XXI. International Young Scientists Conference, Brno, 22. 5. 2019 – lecture, full length contribution. The ability of the immune system to react to stimulants can be measured by functional immunological assays. After stimulation by various mitogens (including pokeweed mitogen, concanavalin A, phytohaemagglutinin and lipopolysaccharide), lymphocytes proliferate and transform into lymphoblasts. The methodology varies between different authors – an optimal sample, mitogen and its concentration, serum type and incubation conditions were determined in this study. Samples obtained from rainbow trout (Oncorhynchus mykiss) included head kidney and peripheral blood collected using different anticoagulants (heparin, EDTA). Mononuclear cells were isolated by density gradient centrifugation. The isolated cells were incubated with different mitogens and foetal bovine serum or trout serum. After the incubation period, labeled thymidine was added to the cell suspension. The amount of thymidine incorporated into DNA was subsequently measured by a liquid scintillation counter in counts per minute (CPM). The highest levels of cell proliferation were observed with peripheral blood lymphocytes (collected using heparin) after 7 days of incubation at 15 °C. Best results were obtained with 50 µg/ml pokeweed mitogen and foetal bovine serum.

Czech name

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Czech description

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Type

O - Miscellaneous

CEP classification

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OECD FORD branch

40301 - Veterinary science

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2019

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů