The aim of the study was to investigate the interaction of lectins with various populations of maturing murine T-lymphocytes, as well as with thymocytes at different stages of apoptosis. Materials and methods. Thymocyte typing of 80 CBA mice was performed by flow cytometry. The binding of lectins to cells in early and late apoptosis induced by the administration of hydrocortisone was also evaluated. Results. The suitability of peanut and Helix pomatia lectins for differentiation of mature and immature mouse thymocytes has been established. 11 lectins bound to living cells, during the transition of cells to the state of early apoptosis, thymocytes were stained with 16 lectins, and upon transition to late apoptosis, 20 of 23 lectins bound to the cells. Conclusion. The use of labeled lectins to assess the stage of murine thymocyte apoptosis does not have obvious advantages over existing methods. The degree of binding of all lectins to thymocytes in apoptosis increases as the charge on the membrane decreases and its permeability increases. For typing thymocytes in the early stages of maturation, peanut and Helix pomatia lectins can be used. Snowdrop and amaryllis lectins are not suitable for differentiation of thymocytes by maturity.
The aim of the study was to compare the ability of various media for cryopreservation of sperm to ensure their viability after thawing and to assess the possibility of using Narcissus pseudonarcissus lectin to determine the viability of native and cryopreserved human sperm by flow cytometry. Materials and methods. Used ejaculate 54 men aged 26 to 47 years, undergoing treatment for infertility. The control was a native ejaculate, which was also used for the in vitro fertilization procedure. Four parallel samples were frozen using various commercial media. After storage and thawing, spermatozoa viability was assessed by flow cytometry using three dyes and Narcissus pseudonarcissus lectin. Results. All assays showed that cryopreservation led to a twofold decrease of sperm viability, dye to the changes in the composition and properties of cell membrane, decrease in mitochondrial membrane potential, as well as the damages of acrosomal complex and nucleus. The lowest decrease in sperm viability was shown for Quinns advantage sperm freezing medium for cryopreservation. Conclusion. Flow cytometry makes it possible to evaluate with high efficiency sperm viability as the part of in vitro fertilization. The results of viability assessment using daffodil lectin make the prediction of in vitro fertilization outcome more accurate.
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