Features of obtaining metaphase chromosomes Caprovinea

Abstract

Obtaining cultures of lymphocyte cells for studying of animal karyotypes is an easily accessible, relatively cheap method and allows obtaining results in a short time. We optimized the usual method with corrected some adjustments to the protocol to obtain metaphase plates of chromosomes of various animal species of good quality. At the same time, such criteria as the use of different media for the cultivation of blood cell cultures and the possibility of using them in different species of animals were taken into account. Blood samples were collected from the jugular vein in goats (n=15), sheep (n=30) and round (n=1). The cell culture was cultivated for 72 h; colchicine was added for 70 h in an amount of 0.5 ?l / ml. The study used nutrient media such as DMEM (Dulbecco?s Modified Eagle?s Medium) и RPMI 1640 (Roswell Park Memorial Institute) and mitogens phytohemagglutinin and Concanavalin A in different concentrations. Optimal parameters for the preparation of metaphase plates with good chromosome distribution were obtained using RPMI 1640 medium supplemented with dry L-glutamine and the simultaneous use of mitogens such as conconavalin and PHA (phytohemagglutinin). The number of plates of good quality when obtaining a culture of lymphocyte cells, depending on the type of animals, ranged from 2.17 to 6.52% in sheep, 33.3-66.7% in goats, and reached 100% in tur. The modified technique made it possible to reduce the loss of cells during the preparation of chromosome preparations. Thus, the results of our study show that the growth and division of lymphocyte cells isolated from the peripheral blood of farm animals depends on the composition of the environment and the species characteristics of the animals.