Blood cells from three different sea turtle species were cultured for approximately 3 weeks in nutrient medium supplemented with recombinant human cytokines known to induce terminal maturation of human hematological stem cells. Cultured turtle erythrocytes were translucent, approximately 10x larger than human erythrocytes, contained a single fluorescent inclusion body, contained nuclear epsilon (embryonic) globin proteins, and, absent of organelles while fresh cells contained few, but well defined mitochondria. Cells with basophilic cytoplasm and in all stages of proliferation were observed in cytokine-supplemented cultures and appeared to possess active protein synthesis.
View Article and Find Full Text PDFBackground: Used leuko-depletion filters (LDFs), containing billions of white blood cells (WBCs), are discarded. Because the steady-state blood contains low quantities of stem and progenitor cells that are retained in LDFs, the viability and the functional properties of mononuclear cells (MNCs) and CD 34+ cells recovered from LDFs were investigated.
Study Design And Methods: WBCs were recovered from LDFs by use of a closed system.
Objective: The aim of this study was to determine the extent of human endogenous retrovirus (HERV) gene translation in megakaryocytes cultured from peripheral blood stem cells of patients with essential thrombocythemia previously reported with platelet-associated HERV sequences and reverse transcriptase activity.
Materials And Methods: Terminally differentiated megakaryocytes derived from circulating stem cells in serum-free medium supplemented with stem cell factor and thrombopoietin were processed for electron microscopic immunostaining using a monoclonal antibody against the gag protein of HERV-K10 and an electron dense gold-labeled secondary antibody.
Results: We found that HERV-K gag protein was detected as clusters in the cytoplasm as well as associated with viral particles budding from the cell membrane and into intracellular vacuoles in megakaryocytes from two patients with essential thrombocythemia.
In earlier studies we identified a putative repressor of the human beta-globin gene, termed beta protein 1 (BP1), which binds to two silencer DNA sequences upstream of the adult human beta-globin gene and to a negative control region upstream of the adult delta-globin gene. Further studies demonstrated an inverse correlation between the binding affinity of the BP1 protein for the distal beta-globin silencer sequence and the severity of sickle cell anemia, suggesting a possible role for BP1 in determining the production of hemoglobin S. We have now cloned a cDNA expressing the BP1 protein.
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