The optimization of the rapid colorimetric assay for the growth and proliferation of peripheral blood lymphocytes
2001
Blahovec, J. | Sobekova, A. (University of Veterinary Medicine, Kosice (Slovak Republic))
There are many different methods for determining cell proliferation. The most frequently used methods are based on the incorporation of radio-labelled nucleotides into DNA. We tried to standardize and optimize one of non-radioactive methods for the determination of bovine peripheral-blood lymphocytes proliferation. The assay is based on the use of yellow methyl tetrazolium salt (MTT) which is cleaved by dehydrogenase activity of active mitochondria to purple formazan. According to our study, the optimal concentration of peripheral-blood lymphocyte is markedly higher than for other commonly-used cell lines. Proliferation of lymphocytes depends on the presence of foetal calf serum and mitogens and the maximum proliferation is been observed after 48 hours of incubation
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