Impact of lymphocyte culture media on the number of metaphases and chromosome band resolution.

P. Chomik,M. Gryzińska,J. Batkowska,G. Jeżewska-Witkowska,J. Kocki

Published 2016 in Postępy Higieny i Medycyny Doświadczalnej

ABSTRACT

A special type of differential staining of chromosomes is replication banding. This staining technique reveals the band pattern characteristic of each homologous pair of chromosomes, which is a reflection of heterogeneous euchromatin structure. Banding enables identification of homologous chromosomes and detection of chromosomal aberrations, both structural and numerical. Slide preparation requires knowledge of many techniques, and the procedure is often different for each laboratory. The aim of the study was to determine the effect of selected media for lymphocyte cultures on the number of metaphases and the band resolution of chromosomes. The study was carried out using cell cultures from whole peripheral blood. The slides were stained by the GTG method. After their removal from the water bath they were immersed in trypsin solution, then rinsed in PBS solution and stained in Giemsa solution. After staining they were rinsed again and left to dry. The study confirmed the effect of selected commercially available cell media on the number of metaphases and band resolution of chromosomes, which have not previously been described. In all of the tests performed, the cell culture, fixation, slide preparation (automatic method), staining, and number of reagents were identical.

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