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Similarly to the production of red blood cells, the production and development of leukocytes takes place in the bone marrow in the process of hematopoiesis. The key stages in the development of leukocytic cell lines are: neutrophilic, eosinophilic and basophilic granulopoiesis, monocytopoiesis, lymphopoiesis and megakaryopoiesis. Leukocytes produced at various developmental stages are characterized by differently shaped nuclei, the presence or absence of nucleoli, and variations in cytoplasm staining. The experiment was carried out on 10 clinically healthy HF cows. Bone marrow aspirate smears stained by the MGG method were analyzed. The study found significant differences in the number, size, and staining intensity of individual myeloid cell lines.
α1,6-fucosyltransferase (FUT8) attaches fucose residues viaan α1,6 linkage to the innermost N-acetylglucosamine residue of N-linked glycans. Glycans with this type of structure are present in GpIIb/GpIIIa complex (CD41a) which is present on megakaryocytes (Mks) and platelets. CD41a is the earliest marker of megakaryocytopoiesis. The aim of this study was to analyse the morphology, phenotype, ploidy level and activity of FUT8 during induced differentiation/maturation of Mk progenitor cells in ex vivoculture. We used SU6656, a selective inhibitor of Src tyrosine kinases, as differentiation-inducing agent for Mks. The addition of SU6656 to the culture system of megakaryocytic progenitors from cord blood CD34+cells and Meg-01 cell line induced their maturation towards later stages of Mk differentiation with increased activity of FUT8. We suggest FUT8 as a candidate for an early marker of differentiation and possibly of the ploidy level of Mks. We confirm a special status of FUT8 in megakaryocytopoiesis.
The aim of the study was an estimation of the histomorphometry of megakaryocytes (MK) in bone marrow in selected myeloproliferative and lymphoproliferative diseases. Bone marrow specimens were obtained by trephine biopsies from 41 patients with polycythaemia rubra vera (PV), idiopathic myelofibrosis (MF), chronic lymphocytic leukaemia (CLL), hairy cell leukaemia (HCL) and diffuse large B-cell lymphoma (L). Morphometric evaluation was performed using a standard program set MicroImage (OLYMPUS). The greatest number of typical nucleated MK, “naked” nuclei, anucleated cytoplasmic fragments and the largest area were found in PV. The circular deviation factor of MK and their nuclei increased in all cases. The greatest number of clusters was observed in PV and HCL. A significant increase in the number of dysplastic and “naked” nuclei of MK was noted in all selected haematological diseases. The presence of neoplastic cells in bone marrow increased the morphological changes in MK. Quantitative and morphometrical significant differentiation of MK in separate microscopic field in the same slides confirms the necessity of performing trephine biopsies in each patient with haematological disorders.
α1,6-Fucosyltransferase (6FucT, E.C. 2.4.1.68) is one of the enzymes involved in the synthesis of N-linked glycans of the GpIIb/IIIa complex (CD41a) which is present on megakaryocytes (MKs) and platelets. In this study, we examined 6FucT activity in ex vivo cultures of immunoselected cord blood CD34+ cells grown in a medium promoting megakaryocytopoiesis. Our results show that the activity of 6FucT increased ahead of, and thereafter concomitantly with, cells expressing the CD41a antigen. When the CD41a+ subpopulation of cells was immunoselected (using anti-CD61 i.e. anti-GpIIIa antibodies), its 6FucT activity increased proportionally to the yield of CD61+ cells. Taking into account the heavy load of 6FucT in platelets and megakaryocytes, we regard this enzyme as a candidate for the earliest marker of MK-commitment in cultured hematopoietic stem cells. Such a marker should allow an earlier detection and earlier transplantation of patients’ own, ex vivo expanded, Mk progenitors.
Plasma membranes of rat platelets produced at normal platelet counts and during early recovery from immune-mediated thrombocytopenia were examined for the contents of carbohydrates, lipids and glycosphingolipids. Glucosylceramide, two monosialo-gangliosides and one disialo-ganglioside were found to be the major glycosphingolipids of platelets. During thrombocytopenia the contents of these glycosphingolipids as well as of ceramides were several fold elevated. Among carbohydrate constituents of platelets and platelet plasma membranes, glycogen content was increased and that of sialic acid decreased. These results are discussed in the light of literature data on relevant biochemical characteristics of megakaryocytes at different stages of maturation and on thrombopoiesis during acute experimental thrombocytopenia.
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