[Isolation and receptor activity of mammalian lymphocytes].

Abstract

The possibilities of lymphocytes from the blood of pigs, cows, sheep and rabbits, using Lymphoprep, Polysep, and Histopaque were investigated. The conditions given by the producer firms were strictly observed when dealing with the separation media, and in cases when the results were unsatisfactory modifications were made with regard to the velocity and duration of centrifugation in carrying out the separation. The functional state of membrane receptors of lymphocytes was checked by rosette tests with erythrocytes for the detection of T-lymphocytes. It was demonstrated that lymphocytes from pig's blood are successfully isolated with all three separating media, when observing the conditions of manipulation as indicated by producer firms. With ruminants the best results were obtained with Lymphoprep. Polysep is not suitable medium for lymphocyte isolation from cow's blood; Histopaque can be used if the separation is carried out at 1880 r. p. m. for 40 minutes. Good results are obtained with Polysep with sheep's blood when the separation is performed at 1880 r. p. m. for 40 minutes. All three media can be used in isolation of lymphocytes from rabbit's blood. The isolation of lymphocytes with Histopaque must be performed by centrifugation at 1520 r. p. m. for the duration of 40 minutes. The separation media investigated do not harm the membrane receptors of lymphocytes.