• Title, Summary, Keyword: megakaryocyte

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Effect of splenectomy on the blood and marrow megakaryocyte picture in dogs (견(犬)에 있어서 비장적출(脾臟摘出)이 혈액(血液) 및 골수거대핵세포상(骨髓巨大核細胞像)에 미치는 영향(影響))

  • Hong, Kyung-tae;Lee, Hyun-beom;Lee, Keun-woo
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.327-336
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    • 1993
  • Present experiments were undertaken in order to clarify the effect of splenectomy on the hematology and marrow megakaryocyte picture and to know the genesis of postsplenectomy thrombocytosis in dogs. Six mongrel dogs weighing 8.5~18㎏ were used, of which three were splenectomized and the other three were laparotomized for comparison. Erythrocyte count, total and differential leukocyte counts, thrombocyte count and packed cell volume measurement were made using the blood samples. In addition, bone marrow samples obtained from the femur at 7th and 23rd day of the operation were examined for the number per low-power field, the diameter, and the distribution frequency of the megakaryocyte. From these experiments, following results were obtained : Erythrocyte count and packed cell volume showed significant decrease beginning on the 15th day of splenectomy. Total and differential leukocyte counts showed marked increase for the first 2 days of postsplenectomy. The thrombocyte count of splenectomized dogs increased from the 2nd day of the operation, reached to the peak count on the 15th day, and returned to the preoperation count by the 28th day. The megakaryocyte count per low-power field of the biopsied preparation increased in according to the increase in thrombocyte count. The megakaryocyte diameter of splenectomized dog showed no increase on the 7th or 23rd day of the operation. However, the distribution frequency of the larger megakaryocyte was higher in the splenectomized dogs than in the laparotomized dogs. The total plasma protein concentration showed no significant change after splenectomy or laparotomy. From these results, it may be concluded that the postsplenectomy thrombocytosis results from the increased megakaryocytopoesis or the activated thrombocytopoesis of the marrow megakaryocytes.

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Acute Megakaryoblastic Leukemia (급성 거핵아구성 백혈병 1례)

  • Kim, Young-Jin;Kim, Tae-Nyun;Hyun, Myung-Soo;Shim, Bong-Sup;Lee, Hyun-Woo;Kim, Jung-Suk
    • Yeungnam University Journal of Medicine
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    • v.8 no.2
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    • pp.209-216
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    • 1991
  • Acute Megakaryoblastic leukemia is a rare and rapidly fatal disease characterized by proliferation of megakaryocyte series and atypical megakaryocytes in the bone marrow. Acute Megakaryoblastic leukemia is suspicious when 1) megakaryocyte in peripheral blood, mixture of large and small mononuclear megakaryoblast in the bone marrow 2) cytoplasmic budding in blast 3) myelofibrosis (dense medullary overgrowth of reticulin fibers) 4) PAS(+), ANAE(+), SBB(-), peroxidase(-) and which is confirmed by platelet peroxidase oxidation on electromicroscope or monoclonal antibody. A case of aute megakaryoblastic leukemia was studied morphologically and monoclonal antibody.

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In Vitro Effect of Interleukin-11 (IL-11) on Megakaryopoiesis from Umbilical Cord Blood Cells (생체 외 제대혈 배양에서 거대핵세포 조혈에 대한 Interleukin-11 (IL-11)의 효과)

  • Lee, Kuk-Kyung;Kim, Chan-Kyu;Lee, Nam-Su;Kim, Sook-Ja;Cheong, Hee-Jeong;Lee, Kyu-Tack;Park, Sung-Kyu;Baick, Seung-Ho;Won, Jong-Ho;Hong, Dae-Sik;Park, Hee-Sook
    • IMMUNE NETWORK
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    • v.3 no.1
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    • pp.47-52
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    • 2003
  • Background: The megakaryopoiesis and platelet production is regulated by several hematopoietc factors such as thrombopoietin (TPO), interleukin-11 (IL-11) and interleukin- 3 (IL-3). IL-11 is a potent stimulator of megakaryopoiesis in vivo, and acts primarily as a megakaryocyte maturation factor in vitro and it can act synergistically with IL-3 and TPO. We performed this study to investigate the effects of recombinant human IL-11 (rhIL-11) with other hematopoietic factors on megakaryocyte colony formation in vitro. Methods: CD34+ cells were separated from umbilical cord blood and megakaryocyte colonies using MegaCult Assay Kit were cultured with rhIL-11, recombinant human IL-3 (rhIL-3), and recombinant human TPO (rhTPO) for 7 and 14 days. The number and percentage of CD34+ and CD41a+ cells were determined by flowcytometry. Results: The number of CD41a+ cells were $0.54{\pm}0.05{\times}10^4$ (rhIL-11 100 ng/ml), $5.32{\pm}0.23{\times}10^4$ (rhIL-3 100 ng/ml), and $8.76{\pm}0.15{\times}10^4$ (rhTPO 50 ng/ml) of total expanded cells during the culture of the purified CD34+ cells in liquid phase for 7 days. The number of CD41a+ cells were increased to $7.47{\pm}0.69{\times}10^4$ (rhIL-3+ rhIL-11), $11.92{\pm}0.19{\times}10^4$ (rhTPO+rhIL-11) of total expanded cells, respectively, during the culture of the purified CD34+ cells in liquid phase for 7 days in the presence of rhIL-11 (100 ng/ml). When the purified CD34+ cells were cultured in semisolid mediaincluding various concentration of rhIL-11, the megakaryocyte colonies were not formed. When the purified CD34+ cells were cultured with rhIL-11 and rhTPO or with rhIL-11 and rhIL-3, the number of megakaryocyte colonies were increased compared with rhTPO or rhIL-3 alone. Conclusion: These results indicate that IL-11 exerts a potent proliferative activity to colony forming unit-megakaryocyte from human umbilical cord blood, and it acts with other hematopoietic factors synergistically.

Production and Characterization of a Monoclonal Antibody against Surface Glycoprotein, gp6 1, on K562 Erythroleukemia Cells (K562 적혈구암 세포주의 표면 당단백질에 대한 단클론항체의 생성 및 특성)

  • 김한도;정재훈;홍선화;김정락;한규형;임운기;유미애;이경희;강호성
    • The Korean Journal of Zoology
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    • v.39 no.1
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    • pp.12-20
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    • 1996
  • A multipotential hematopojetic cell line, 1(562 cell, was differentiated into megakaryocyte by a chemical inducer, PMA, with an enhanced expression of gpIlla accompaning with a distinct morphological change. On the other hand, 1(562 cells were differentiated into erythrocytes by other chemical inducers, DMSO or butyrate, with a concomitant increase in hemoglobin accumulation. An antigen of apparent molecular weight of 61 kDa was identified on the surface of 1(562 cells by using monoclonal antibody raised against 1(562 cells. The antigen was considered to be a glycoprotein molecule rich in sialic acids and the epitope of antigen was sensitive to neuraminidase digestion or peroxidase oxidation, but resistant to heat treatment. The 61 kDa surface antigen was increased or decreased in its expression along differentiation of 1(562 cells into megakaryocytes or erythrocytes, respedively.

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Megakaryocyte Colony Formation of Fetal Liver Cells (태아 간세포의 거핵구 집락형성)

  • Kwon, Byung O;Ju, Hye Young;Kim, Chun Soo;Jeon, Dong Seok;Kim, Jong In;Kim, Heung Sik
    • Clinical and Experimental Pediatrics
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    • v.45 no.2
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    • pp.247-255
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    • 2002
  • Purpose : This study was undertaken to obtain basic data about the megakaryocyte colony formation of fetal liver cells by using immunocytochemical staining and ex vivo culture with growth factors. Methods : The mononuclear cells were isolated from fetal liver and bone marrow with idiopathic thrombocytopenic purpura(ITP) and pancytopenia. These mononuclear cells were cultured in $MegaCult^{TM}-C$(Stem Cell Tech, Canada) media in the presence of growth factors and CFU-Megakaryocyte( CFU-Mk) colonies were counted on day 12. The expansion of CD34+ and CD41+ cell was analyzed by flow cytometry after 5 days incubation using flask culture. Results : The numbers of CFU-Mk colonies of mononuclear cells obtained from fetal liver in the 11th week gestational age were more than those in the 19th week specimens; growth factors could not enhance the colony expansion in all cases. Total numbers of CFU-Mk colony of fetal liver cells were higher than bone marrow from ITP or pancytopenia groups. The numbers of pure or large CFU-Mk colonies of fetal liver cells were also higher than bone marrow specimens. The rate of CD34+ cell expression of fetal liver was increased after flask culture and the enhancement effect of epression was seen only in cases which added thrombopoietin. The rate of CD41+ cell expression of fetal liver was increased after incubation, but the enhancement effect of growth factors was unclear. Conclusion : This study revealed good results about the megakaryocyte colony assay of fetal liver mononuclear cells using $MegaCult^{TM}-C$ media. This study suggests that the fetal liver could be a good source of megakaryocytic progenitor cells for clinical application in hematopoietic stem cell transplantation.

Gene Expression Analysis of Megakaryocytes Derived from Human Umbilical Cord $CD34^+$ Cells by Thrombopoietin

  • Kim, Jeong-Ah;Kim, Hyung-Lae
    • Genomics & Informatics
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    • v.3 no.1
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    • pp.8-14
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    • 2005
  • Although much is known about the molecular biology of platelets, the megakaryocytes' (MKs) molecular biology was not understood so well because of their rareness. By the cloning and characterization of thrombopoietin (TPO), which is the principal regulator of the growth and development of the MKs, researches on the MKs have been growing rapidly. To understand megakaryocytopoiesis, we investigated the gene expression profile of the MKs using oligonucleotide microarray where 10,108 unique genes were spotted. Comparing the fluorescence intensities of which ratio is $\ge$ ${\mid}2{\mid}$, 372 genes were up-regulated and 541 genes were down-regulated in MKs. For confirmatory expression, RNase protection assay (RPA) establishing abundant apoptotic gene expression was carried out. In MKs, many of the known genes, including several platelet related genes, GATA binding protein were highly expressed. Particularly, TGF beta, clusterin (complement lysis inhibitor), and thymosin beta 4 (actin-sequestering molecules) were expressed highly in MKs. As MKs specific expressed genes may regulate normal and pathologic platelet (and/or MK) functions, the transcript profiling using microarray was useful on molecular understanding of MKs,

A Study on Radio-Protection Mechanism of Platelet Cells After Injection of Alliin (알리인 투여 후 혈소판의 방사선 방어기전 연구)

  • Ji, Tae-Jeong
    • Journal of radiological science and technology
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    • v.33 no.3
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    • pp.185-192
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    • 2010
  • Platelets originating from Megakaryocyte are sensitive to radiation along with white blood cells, and thus these platelets are used as an index of radiation hazard as they decrease in advance. Thus, when there is a scarcity of platelets, dot hemorrhage occurs and it leads to decrease of blood corpuscle and a decline in immunity. In particular, when 4~6 Gy whole body irradiation is received, after three weeks, the platelets will decrease to the lowest level, which can be a cause of death by bleeding and anemia. Therefore, this study tried to identify the mechanism of platelet damage and protection effect. The protection substance used in the experiment is Alliin, which is a component of garlic, and it was observed by an Transmission Electron Microscope(TEM) after its injection to the rat's tail vein. In the study, it was found that the cell membrane was severely damaged in a 10-day progressed platelet organ after receiving 5 Gy irradiation. It billowed as balloon-like figure and the glycocalyx became hyperplasia. The minute organ was damaged to the point that it was beyond recognition in a 20-day progressed platelet organ after receiving irradiation, and the cytoplasmic contents were exposed to epilepsy parts and outrageously damaged. Furthermore, the form of granules could also not be observed. A hole was formed in the middle, and the damaged organ was found in a 30-day progressed platelet. However, the form of granules was consistently maintained in the experiment group injecting Alliin, as with the control group, and there was no damage to the cell membrane recognized. Thus, it was possible to verify the effectiveness of radiation protection of the platelet when Alliin was injected to the blood vessel.

Ex vivo Expansion and Clonal Maintenance of CD34+ Selected Cells from Cord Blood and Peripheral Blood (제대혈 및 말포혈로부터 분리한 CD34 양성 세포의 체외 증폭 및 클론 유지)

  • Kim, Soon Ki;Ghil, Hye Yoon;Song, Sun U.;Choi, Jong Weon;Park, Sang Kyu
    • Clinical and Experimental Pediatrics
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    • v.48 no.8
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    • pp.894-900
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    • 2005
  • Purpose : Because of the unavailability of marrow transplantation, umbilical cord blood (CB) is increasingly being used. We evaluated the potential of ex vivo expansion and clonality in CD34+ cells separated from cord blood source and mobilized peripheral blood (PB) in a serum-free media. Methods : The CD34+ cells, selected from CB and mobilized PB, were expanded with hematopoietic growth factors. They were then cultured for burst-forming units of erythrocytes (BFU-E), colony-forming units of granulocytes and monocytes (CFU-GM) and colony-forming units of megakaryocytes (CFU-Mk) at culture days 0, day 4, day 7, and day 14 with various growth factors. Results : The CB-selected CD34+ cells showed significantly higher total cell expansion than those from the PB at day 7 (2 fold increase than PB). The CB-selected CD34+ cells produced more BFU-E colonies than did the PB on culture at days 7 and at day 14. Also, the CB-selected CD34+ cells produced more CFU-Mk colonies than did the PB on culture at day 4 and at day 7. Conclusion : The ex vivo expansion of the CB cells may be promising in producing total cellular expansion, CFU-Mk and BFU-E compared with PB for 7 to 14 days. The growth factors combination including megakaryocyte growth and development, flt3-ligand and interleukin-3 showed more expansion in the view of total cells and clonal maintenance compared with less combination.

2-(trimethylammonium)ethyl (R)-3-methoxy-3-oxo-2-stearamidopropyl phosphate enhances thrombopoietin-induced megakaryocytic differentiation and plateletogenesis

  • Kim, Jusong;Jin, Guanghai;Lee, Jisu;Lee, Kyeong;Bae, Yun Soo;Kim, Jaesang
    • BMB Reports
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    • v.52 no.7
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    • pp.434-438
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    • 2019
  • We have previously reported the effects of 2-(trimethylammonium)ethyl (R)-3-methoxy-3-oxo-2-stearamidopropyl phosphate [(R)-TEMOSPho], a synthetic phospholipid, on megakaryocytic differentiation of myeloid leukemia cells. Here, we demonstrate that (R)-TEMOSPho enhances megakaryopoiesis and plateletogenesis from primary hematopoietic stem cells (HSCs) induced by thrombopoietin (TPO). Specifically, we demonstrate at sub-saturation levels of TPO, the addition of (R)-TEMOSPho enhances differentiation and maturation of megakaryocytes (MKs) from murine HSCs derived from fetal liver. Furthermore, we show that production of platelets with (R)-TEMOSPho in combination with TPO is also more efficient than TPO alone and that platelets generated in vitro with these two agents are as functional as those from TPO alone. TPO can thus be partly replaced by or supplemented with (R)-TEMOSPho, and this in turn implies that (R)-TEMOSPho can be useful in efficient platelet production in vitro and potentially be a valuable option in designing cell-based therapy.