DOCSLIB.ORG

Establishment of a Human Megakaryoblastic Cell Line (T-33) from Chronic Myelogenous Leukemia in Megakaryoblastic Crisis1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Establishment of a Human Megakaryoblastic Cell Line (T-33) from Chronic Myelogenous Leukemia in Megakaryoblastic Crisis1 [CANCER RESEARCH 48. 6137-6144, November 1. 1988] Establishment of a Human Megakaryoblastic Cell Line (T-33) from Chronic Myelogenous Leukemia in Megakaryoblastic Crisis1 Tsuyoshi Tange,2 Kazuhiko Nakahara, Kinuko Mitani, leharu Yamasaki, Hideyuki Yasuda, Fumihiko Tanaka, Masaru Mizuguchi, Hideaki Oda, Yutaka Yatomi, Riichiro Takanashi, Seitoku Fujioka, Kazuyoshi Yamaguchi, and Yoshinori Urano Department of Pathology and the First Department of Internal Medicine, University of Tokyo, and Mitsui Memorial Hospital, Tokyo, Japan ABSTRACT peripheral blood of a female with megakaryoblastic crisis of CML (13) and here report this cell line termed as T-33 as a A megakaryoblastic cell line, termed T-33, was established from the human megakaryoblastic cell line capable of differentiating into peripheral blood of a patient with Philadelphia chromosome-positive chronic myelogenous leukemia in megakaryoblastic crisis. T-33 cells megakaryocyte. have been maintained in RPMI 1640 medium containing 10% fetal calf serum in a single cell suspension with a doubling time of 24-36 h for MATERIALS AND METHODS over 2 years. Giemsa-banded karyotypes were female hyperdiploid with Case History. A 55-year-old female was diagnosed as Ph'-positive a modal chromosomal number of 51, all cells including Philadelphia chromosome. The cells showed strong positivity for periodic acid-Schiff CML and treated with busulfan at Mitsui Memorial Hospital (Tokyo) and a-naphthyl acetate esterase, and weak for a-naphthyl butyrate in 1978. In 1983, blastic crisis of CML was suspected because of the esterase, but were negative for myeloperoxidase. Flow cytometric analysis appearance of hip joint pain but the bone marrow was still in the of cell surface markers showed the existence of HLA-DR, MY-7, MY- chronic phase with myelofibrosis. Thereafter, vincristine, prednisolone, 9, and a platelet antigen (Yukb), and no markers for T- or B-lymphocytes. and 6-mercaptopurine were given when the leucocyte count increased. Most of the cells fixed with acetone were positive for Factor VIII, platelet On the last admission in August, 1985, the leucocyte count was 13,200 glycoprotein Hb-IIIa, Ilia (Yukb), and Ib, but negative for glycophorin A (blasts, 2.5%) and the bone marrow was dry tap, the biopsy revealing osteomyelofibrosis. The patient became refractory' to any drugs and the and hemoglobin. Ultrastructural platelet peroxidase was demonstrated in 2-3% of cells and the percentage of positive cells increased up to 20% leucocyte count increased up to 26,700 (blasts, 15.5%). Then, ultra- after the treatment with 12-O-tetradecanoylphorbol-13-acetate. The cells structural study of the peripheral blood blasts revealed the positivity contained small dense granules negative for platelet peroxidase, their for platelet peroxidase. leading to a diagnosis of megakaryoblastic crisis number increasing threefold after 12-0-tetradecanoylphorbol-13-acetate of CML. The patient died of pulmonary edema on October 30, 1985. Cell Culture. On October 23, 1985, 10 ml of heparinized peripheral treatment. Such treated cells frequently showed a complex of the demar blood was obtained, when the white blood cell count was 5000/mm3 cation membrane in the cytoplasm. T-33 responded thrombin to exhibit calcium influx. This cell line may be useful for the study of the early (blasts, 50%). The sample was diluted in 0.1 mol PBS. pH 7.3, and the mononuclear cells were separated by Ficoll-Conray gradient centrifu- stage of megakaryocytic differentiation in human megakaryopoiesis. gation. After washing three times in PBS, cells were cultured at a concentration of 106/ml in Falcon plastic tissue culture bottles contain INTRODUCTION ing 7 ml of RPMI 1640 medium (GIBCO, Grand Island, NY) and 10% PCS (Flow Laboratories, North Ryde, NSW, Australia) with penicillin Since it became possible to identify a megakaryoblast by ( 100 U/ml) and streptomycin (50 Mg/ml) at 37°C,5%CO2. The cultured demonstrating ultrastructural PRO,3 platelet GP Ib and Hb- cells were fed once or twice weekly by half medium change. Illa, and coagulant factors such as factor VIII or IV (1-4), an Morphological Observation. Cultured cells were observed by the increased number of cases with megakaryoblastic leukemia or Nikon inverted-phase contrast microscope (Nikon, Tokyo). The cells megakaryoblastic crisis of CML have been reported (2, 5-7). were smeared on the slide glass and stained with May-Gruenwald- Giemsa or Wright's staining. Recently, a few megakaryoblastic cell lines such as MEG-01 have been established (8, 9) but MEG-01 did not morphologi Chromosomal Analysis. In absence of Colcemid, the cultured cells were treated with 75 mmol/liter KC1 hypotonie solution for 30 min at cally show any evident megakaryocytic differentiation (8). Cell 37°C,andfixed with methanol-acetic acid solution. Chromosomes were lines established by Morgan et al. did not exhibit any granules banded by the trypsin-Giemsa method. in the cytoplasm in spite of the development of a megakaryocy Cytochemistry. The cultured cells suspended in 10% bovine serum tic membrane system (9). A few megakaryocytic cell lines such albumin were smeared on the slide glass, air dried, and fixed by the use as EST-IU and CHRF-288 have also been reported (10-12). of published methods for cytochemical stainings. EST-IU seems to lack the DM, while CHRF-288 is a mega Antibodies. Most of the monoclonal antibodies used were purchased karyocytic tumor cell line forming a s.c. tumor in nude mice, from Becton-Dickinson Co. (Sunnyvale, CA) except MY-4 (CD 14), but nonleukemic. We have established a human cell line releas MY-7 (CD 13), and MY-9 (CD 33) which were from Coulter Co. ing a virus-like particle into the culture supernatant from the (Hialeah, FL). TP-80 (14), TP-82 (15). and TM-83, murine IgG mono clonal antibodies specific for human platelet GP Ilb-IIIa were kindly Received 12/28/87; revised 6/25/88; accepted 7/26/88. provided, TP-80 and TP-82 were by Dr. Hiroo Maeda (University of The costs of publication of this article were defrayed in part by the payment Tokyo Hospital), and TM-83 was by Dr. Kenjiro Tanoue (Tokyo of page charges. This article must therefore be hereby marked advertisement in Metropolitan Institute of Medical Science, Tokyo). Anti-Yukh. kindly accordance with 18 U.S.C. Section 1734 solely to indicate this fact. ' Supported in part by Grant-in-Aids for Scientific Research (61304036) from provided by Dr. Yoichi Shibata (Toranomon Hospital, Tokyo), is a the Ministry of Education and for Cancer Research (61-11) from the Ministry of maternal polyvalent antibody, human IgG, reacted to fetal platelets. Health and Welfare in Japan. Immunofluorescence Technique. For direct staining, 5 x IO5of T-33 2To whom requests for reprints should be addressed, at the Department of were suspended in 20 ^1of each FITC-conjugated monoclonal antibody Pathology, Faculty of Medicine, the University of Tokyo, 7-3-1, Hongo. Bunkyo- (anti-Leu-1, -2, -3, -4, -7, -10, -11, -12, -Ml, and -M3, anti-HLA-DR, ku. Tokyo 113. Japan. 1The abbreviations used are: PPO, platelet peroxidase; GP. glycoprotein; anti-CALLA, or anti-IL-2) and incubated on ice for 30 min. The cells CML, chronic myelogenous leukemia: Ph', Philadelphia chromosome: NAE, a- were then washed three times with PBS containing 2% fetal calf serum naphthyl acetate esterase; NBE, a-butyrate esterase; DM, demarcation membrane; (PBS-FCS) and resuspended in 1 ml of PBS-FCS. PBS, phosphate buffered saline; PCS, fetal calf serum; FITC, fluorescein isothi- ocyanate; PAP, peroxidase-anti-peroxidase;TPA. 12-O-tetradecanoylphorbol-13- For indirect staining, the same number of T-33 were suspended in acetate. 20 M' of a nonlabeled antibody (MY-4, MY-7, MY-9, TP-80, TP-82, 6137 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1988 American Association for Cancer Research. or Yukb) and incubated on ice for 30 min. After washing three times with PBS-FCS. the cells were resuspended in 20 ^1 of appropriately diluted FITC-conjugated goat anti-mouse immunoglobulins (Cappel; Copper Biochemical, Inc., Malvenu PA) or FITC-conjugated rabbit anti-human immunoglobulins (kindly provided by Dr. Ko Okumura, Juntendo University School of Medicine, Tokyo), followed by incuba tion for 30 min on ice. Then, the cells were washed three times and suspended in 1 ml of PBS-FCS. Cytofluorographic analysis was per formed by a fluorescence-activated cell sorter, FACS-420 (Becton Dick inson, Sunnyvale, CA). Immunostaining of Intracytoplasmic Antigens. After the cells were washed three times, suspended in 10% bovine serum albumin solution, smeared on slides, and fixed with acetone, cytoplasmic antigens such as platelet GP (Ilb-IIIa, Illa, Ib), Factor VIII, hemoglobin A, and glycophorin A were stained with the indirect PAP immunostaining method using monoclonal or polyvalent antibodies. Scanning Electron Microscopy. Cells in the culture bottle were fixed with 1% glutaraldehyde for 30 min and postfixed with 0.5% osmium tetroxide for 20 min. Fixed cells were dehydrated in a graded series of ethanol and coated with gold. Samples were observed by a scanning electron microscope (Hitachi S450LB). Transmission Electron Microscopy. Cultured cells were centrifuged at 3000-5000 rpm for 10 min by Eppendorfs centrifuge and the pelleted cells were fixed in 2.5% glutaraldehyde for 2 h at 4°Cand postfixed in 1% osmium tetroxide for 1 h. Fixed cells were dehydrated in a graded series of ethanol and embedded in EPON. Ultrathin B sectioning was performed with a diamond cutter and sections were stained with uranyl acetate and lead citrate, and observed by JEM 100C electron microscopy (JEM, Tokyo). Ultrastructural Platelet Peroxidase (PPO). Cells unfixed or fixed with periodate-lysine-paraformaldehyde fixative solution were incu bated in 3,3-diaminobenzidine tetrahydrochloride solution according to the method previously described (2). Then, the procedure was proc essed in the same way as described above for the transmission electron microscopy.
Load more

Recommended publications
  • Development of Megakaryoblastic Leukaemia in Runx1-Evi1 Knock-In Chimaeric Mouse
    Letter to the Editor 1458 control. The Mcl-1-specific T-cell clone did not kill this cell line Ms. Bodil K. Jakobsen, Department of Clinical Immunology, (Figure 1c). University Hospital, Copenhagen, for HLA-typing of patient blood The lower rates of relapse in allogeneic transplantation samples. This study was supported by grants from the Danish compared with autologous bone marrow transplantation, the Medical Research Council, The Novo Nordisk Foundation, The striking clinical benefit of donor-lymphocyte infusions as well as Danish Cancer Society, The John and Birthe Meyer Foundation, the finding that human T cells can destroy chemotherapy- and Danish Cancer Research Foundation. resistant cell lines from chronic myeloid leukemia and multiple RB Sørensen1, OJ Nielsen2, P thor Straten1 and MH Andersen1 myeloma, have prompted development of immunotherapeutic 1 strategies against hematological cancers.3 Among these ap- Tumor Immunology Group, Institute of Cancer Biology, Danish Cancer Society, Copenhagen, Denmark and proaches, active specific immunization or vaccination is 2Department of Hematology, State University Hospital, emerging as a valuable tool to boost the adaptive immune Copenhagen, Denmark. system against malignant cells. In this regard, the identification E-mail: [email protected] of leukemia-associated antigens is crucial. However, very few antigens are characterized in a conceptual framework in which the biology, microenvironment, and conventional disease management have been taken into consideration. Myeloid cell References factor-1 (Mcl-1) is a death-inhibiting member of the Bcl-2 family that is expressed in early monocyte differentiation. Elevated 1 Andersen MH, Becker JC, thor Straten P. The anti-apoptotic member levels of Mcl-1 have been reported for a number of solid and of the Bcl-2 family Mcl-1 is a CTL target in cancer patients.
    [Show full text]
  • Erythroid and Megakaryocytic Transformation
    Oncogene (2007) 26, 6803–6815 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc REVIEW Erythroid and megakaryocytic transformation A Wickrema1 and JD Crispino2 1Section of Hematology/Oncology, University of Chicago, Chicago, IL, USA and 2Division of Hematology/Oncology, Northwestern University, Chicago, IL, USA Red blood cells and megakaryocytes arise from a common Accumulated evidence mostly from studies with precursor, the megakaryocyte-erythroid progenitor and mouse models and human primary cells suggests that share many regulators including the transcription factors cellular expansion and differentiation occur concur- GATA-1 and GFI-1B and signaling molecules such as JAK2 rently until the late stages of erythroid differentiation and STAT5. These lineages also share the distinction (polychromatic/orthochromatic) at which point the cells of being associated with rare, but aggressive malignancies exit the cell cycle and undergo terminal maturation that have very poor prognoses. In this review, we (Wickrema et al., 1992; Ney and D’Andrea, 2000; will briefly summarize features of normal development of Koury et al., 2002). A disruption of the balance between red blood cells and megakaryocytes and also highlight erythroid cell expansion and differentiation results in events that lead to their leukemic transformation. It is either myeloproliferative disorders (MPDs) such as clear that much more work needs to be done to improve our polycythemia vera, myelodysplastic syndrome, or rarely understanding of the unique biology of these leukemias in erythroleukemia. Furthermore, some patients initially and to pave the way for novel targeted therapeutics. diagnosed with MPDs ultimately progress to erythro- Oncogene (2007) 26, 6803–6815; doi:10.1038/sj.onc.1210763 leukemia.
    [Show full text]
  • Exposure of Patient-Derived Mesenchymal Stromal Cells To
    Author Manuscript Published OnlineFirst on July 8, 2020; DOI: 10.1158/1541-7786.MCR-20-0091 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. 1 Research Article 2 Exposure of patient-derived mesenchymal stromal cells to TGFB1 supports fibrosis 3 induction in a pediatric acute megakaryoblastic leukemia model 4 Theresa Hack1, Stefanie Bertram2, Helen Blair3, Verena Börger4, Guntram Büsche5, Lora 5 Denson1, Enrico Fruth1, Bernd Giebel4, Olaf Heidenreich3,6, Ludger Klein-Hitpass7, 6 Laxmikanth Kollipara8, Stephanie Sendker1, Albert Sickmann8,9,10, Christiane Walter1, Nils 7 von Neuhoff1, Helmut Hanenberg1,11, Dirk Reinhardt1, Markus Schneider1,* and Mareike 8 Rasche1,* 9 1 Department of Pediatric Hematology and Oncology, University Children’s Hospital Essen, 10 Essen, Germany 11 2 Department of Pathology, University Hospital Essen, Essen, Germany 12 3 Newcastle University, Wolfson Childhood Cancer Research Centre, Translation and Clinical 13 Research Institute, Newcastle upon Tyne, United Kingdom 14 4 Institute for Transfusion Medicine, University Hospital Essen, Essen, Germany 15 5 Department of Pathology, Hannover Medical School, Hannover, Germany 16 6 Princess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands 17 7 Department of Cell Biology, University Hospital Essen, Essen, Germany 18 8 Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V., Dortmund, Germany 19 9 Department of Chemistry, College of Physical Sciences, University of Aberdeen, Aberdeen, 20 Scotland, United Kingdom 21 10 Medizinische Fakultät, Medizinische Proteom-Center (MPC), Ruhr-Universität Bochum, 22 Bochum, Germany 23 11 Department of Otorhinolaryngology and Head/Neck Surgery, Heinrich Heine University, 24 Düsseldorf, Germany 25 * These authors contributed equally to this work.
    [Show full text]
  • Bleeding Fevers! Thrombocytopenia and Neutropenia
    Bleeding fevers! Thrombocytopenia and neutropenia Faculty of Physician Associates 4th National CPD Conference Monday 21st October 2019, Royal College of Physicians, London @jasaunders90 | #FPAConf19 Jamie Saunders MSc PA-R Physician Associate in Haematology, Guy’s and St Thomas’ NHS Foundation Trust Board Member, Faculty of Physician Associates Bleeding fevers; Thrombocytopenia and neutropenia Disclosures / Conflicts of interest Nothing to declare Professional Affiliations Board Member, Faculty of Physician Associates Communication Committee, British Society for Haematology Education Committee, British Society for Haematology Bleeding fevers; Thrombocytopenia and neutropenia What’s going to be covered? - Thrombocytopenia (low platelets) - Neutropenia (low neutrophils) Bleeding fevers; Thrombocytopenia and neutropenia Thrombocytopenia Bleeding fevers; Thrombocytopenia (low platelets) Pluripotent Haematopoietic Stem Cell Myeloid Stem Cell Lymphoid Stem Cell A load of random cells Lymphoblast B-Cell Progenitor Natural Killer (NK) Precursor Megakaryoblast Proerythroblast Myeloblast T-Cell Progenitor Reticulocyte Megakaryocyte Promyelocyte Mature B-Cell Myelocyte NK-Cell Platelets Red blood cells T-Cell Metamyelocyte IgM Antibody Plasma Cell Secreting B-Cell Basophil Neutrophil Eosinophil IgE, IgG, IgA IgM antibodies antibodies Bleeding fevers; Thrombocytopenia (low platelets) Platelet physiology Mega Liver TPO (Thrombopoietin) TPO-receptor No negative feedback to liver Plt Bleeding fevers; Thrombocytopenia (low platelets) Platelet physiology
    [Show full text]
  • Pre and Postnatal Hematopoiesis
    Pre_ and postnatal hematopoiesis Assoc. Prof. Sinan Özkavukcu Department of Histology and Embryology Lab Director, Center for Assisted Reproduction, Dep. of Obstetrics and Gynecology [email protected] 3 8 6 40 8 28 18 E Hemopoiesis (Hematopoiesis) • It is carried out in hematopoietic organs. • Erythropoiesis • Leukopoiesis • Thrombopoiesis ■Erythrocytes, platelets and granulocytes (neutrophils, eosinophils, basophil leukocytes) of blood cells are produced in myeloreticular tissue (red bone marrow). ■Agranulocytes (lymphocytes and monocytes); they are made both in the red bone marrow and in the lymphoreticular tissues (lymphoid organs). Ensuring continuity • The circulating blood cells have certain lifetimes. The cells are constantly destroyed and renewed. Therefore, a continuous production dynamics is needed. Blood product Life span Red blood cells 120 days Fetal red blood cells 90 days Platelets 7-12 days Transfused platelets 36 hours 8-12 hours in circulation Neutrophils 4-5 days in tissue Prenatal hematopoiesis • Yolk sac Stage 3rd Week Hemangioblast formation Prenatal Hemopoez Mesoblastic phase (2nd week-mesoderm of the yolk sac) Hepatosplenothymic phase Liver (6th week) Spleen (8th week) Thymus (8th week) Medullalymphatic phase (3-5th month) Temporary blood islets of the yolk sac • In the 3rd week of embryological development, mesodermal cells in the yolk sac wall are differentiated into hemangioblast cells. • These cells are the precursors of both blood cells and endothelial cells that will form the vascular system. • Blood precursors formed in this region are temporary. • The main hematopoietic stem cells develop from the mesoderm surrounding the aorta, called the aorta-gonad-mesonephros region (AGM), next to the developing mesonephric kidney. • These cells colonize the liver and form the main fetal hematopoietic organ (2-7th month of pregnancy) • Cells in the liver then settle into the bone marrow, and from the 7th month of pregnancy, the bone marrow becomes the final production center 1.
    [Show full text]
  • Hematopoyesis
    HEMATOPOYESIS Dr Blaz Lesina Otoño de 2009 2.5 billones de Gr Kg/peso/día 2.5 billones de Plq Kg/peso/día 1 billon de Gran Kg/peso/día Esqueleto 600 gr de tejido hematopoyético 70% pelvis, vértebras y esternón Saco vitelino Médula ósea 100 Hígado Vértebra 80 Esternón 60 40 Bazo Tibia Costilla ACTIVIDAD ACTIVIDAD HEMATOPOYETICA 20 Femur 1 2 3 4 5 6 7 8 9 10 20 30 40 50 60 70 80 90 Meses de gestación Edad en años Nacimiento Routes a stem cell can take self-renew differentiate Hematopoietic Cell Differentiation CFU-L pluripotential blast colony-forming cell self renew lymphocyte CFU-GEMM self renew BFU-E CFU-GM CFU-Eo CFU-Bas CFU-Meg CFU-E CFU-G CFU-M erythrocyte neutrophil monocyte eosinophil basophil megakaryocyte Plasticidad Célula troncal Tejido Tejido no Referencias convencional convencional Hematopoyética Células de la Nervioso epitelial Petersen 1999, sangre hepático músculo Krause 2001 esquelético y cardiaco Neuronal Nervioso Sangre músculo Bjornson1999, Galli 2000 Muscular Muscular Nervioso sangre Gussoni 1999, Jackson 1999 Mesenquimática Óseo, cartílago Koppen 1999, Ito adiposo músculo 2001 mervioso pulmón estroma hematopoyético renal Dale, D. C. et al. Blood 2008;112:935-945 Lodish y cols.: ”Molecular Cell Biology”.5º Ed. W.H. Freeman 2004 CD34 +, CD34 +, CD38 ++, CD38 --, CD38 -- CD3, cc--kitkit + (CD117) cc--kitkit ++ CD4, CD8,……. Hematopoietic Microenvironment Stromal cells : fibroblasts endothelial cells adipocytes Growth Factors Basal Hematopoiesis SCF IL-6 GM-CSF G-CSF SCF: stem cell factor GM-CSF: granulocyte-macrophage colony-stimulating factor G-CSF: granulocyte colony-stimulating factor IL-1 TNF ααα Ag SCF GM-CSF IL-1 G-CSF IL-3 IL-6 GM-CSF Ag IL-4 TNF ααα Papayannopoulou, T.
    [Show full text]
  • Reticulated Platelets: Analytical Aspects and Clinical Utility
    DOI 10.1515/cclm-2014-0165 Clin Chem Lab Med 2014; 52(8): 1107–1117 Review Johannes J.M.L. Hoffmann* Reticulated platelets: analytical aspects and clinical utility Abstract: Reticulated platelets are immature platelets Formation and maturation circulating in blood; they reflect the activity of megakary- opoiesis in the bone marrow. Therefore, they can be used of megakaryocytes as a non-invasive test in patients with thrombocytopenia in various clinical conditions. The preferred method of The study of blood platelets or thrombocytes started in analysis is by flow cytometry. However, there is an evident the late 19th century, when Wright had developed his lack of analytical standardization, making it difficult to variant of the Romanovsky stain that allowed detailed compare results obtained in different laboratories. Cur- observations of the smallest blood elements [1]. Plate- rently, two types of hematology analyzers are on the mar- lets represent the terminal stage of megakaryopoiesis, ket offering fully automated measurement of reticulated a highly complex sequence of events that can be traced or immature platelets: the high end analyzers manufac- back to the generation of the pluripotent hematopoietic tured by Sysmex (XE- and XN-series) and Abbott (CELL- stem cell in bone marrow. This pluripotent stem cell pro- DYN Sapphire). Although the methods are essentially liferates and differentiates via several intermediates into different and cannot be used interchangeably, both have a megakaryoblast and eventually into a megakaryocyte been proven to have clinical utility. Reticulated or imma- (Figure 1). Megakaryopoiesis is regulated by various ture platelet assays are useful for the differential diagno- growth factors and cytokines, of which thrombopoietin sis of thrombocytopenia and for monitoring bone marrow (TPO) is the most important.
    [Show full text]
  • Immunology-Sheet-2.Pdf
    2 Obada Froukh Obada Froukh Obada Froukh Anas Abu-Humaidan Immunology We will talk about the cells of the immune system. Immune system has cells, molecules and physical barrier which do the immunity job. •The cells of the innate and adaptive immune system are normally present as circulating cells in the blood and lymph, as anatomically defined collections in lymphoid organs, and as scattered cells in virtually all tissues. The majority of the immune cells and the cells in the blood come from the bone marrow, so in the bone marrow we have stem cells (Hematopoietic stem cell) that give other stem cells: 1) Lymphoid stem cells: Differentiate into Natural killer cell (NK cells), T-cell and B-cell (T and B cells are in the adaptive immunity and we will talk about them in the next lecture). 2) Myeloid stem cells (MSC): which differentiate into the majority of the cells in the blood, some of these cells have a relation with the immune system and others don’t have a relation with the Immune system. A) Cells that don’t have a relation with immune system: 1- Erythroblast: is differentiated into Erythrocytes that will lose its nucleus and is converted into (RBCs). 2-Megakaryoblast: is differentiated into Megakaryocytes which is converted into platelets after fragmentation. B) Cells that have a relation with immune system: 1) Putative mast cell precursor: scientists know that the mast cell has a precursor, but we don’t know from where this precursor is originated and hence it is putative. 2) Myeloblast: It gives us three precursor cells that will differentiate into three cells known together as Granulocytes (because all of them have granules).
    [Show full text]
  • UNIT-I Topic- Hematopoiesis Hematopoiesis
    FACULTY OF ENGINEERING & TECHNOLOGY TOPIC UNIT-I Topic- Hematopoiesis Hematopoiesis Production of red blood cells, white cells, and platelets (blood cell formation) Anatomy of Hematopoiesis: .First Trimester: Yolk Sac .Second Trimester: Liver and Spleen .Third Trimester: Central, Peripheral Skeleton . Adulthood: Axial Skeleton, Vertebral Bodies, Sternum, Ribs, Pelvis. Hematopoiesis may re-expand into fetal sites in times of severe demand, e.g. thalassemia, MF (Extramedullary hematopoiesis). The cellular pathways of Hematopoiesis: •Overall Cellular Organization of Hematopoiesis: 1. Stem Cells: o Totipotential. o Multipotential. 2. Progenitor Cells. 3. Precursor Cells. 4. Effector Cells. The cellular pathways of hematopoiesis: I. Stem cells: Totipotential Stem Cells :"The Godfather Cells”: 1. Unlimited Self-Renewal. 2. Unlimited Differentiation: can give rise to any cellular element. 3. Present in Marrow in Small Numbers. 4. Highly Resistant to Chemotherapy. 5. Reside Primarily in Marrow. Small numbers circulate in blood. 6. Surface Antigen -CD34+ ("Cluster of Differentiation 34"). 7. Look like Small Lymphocytes. 8. Dysfunction leads to aplastic anemia or certain types of leukemia. Multipotential Stem Cells: -i.e. Lymphoid, Myeloid Stem Cells 1. Derived from Totipotential Stem Cells. 2. Capable of Extended Self Renewal. 3. Capable of Extended Differentiation • Lymphoid stem cells give rise to all categories of mature lymphocytes. • Myeloid stem cells give rise to red cells, granulocytes, monocytes, and platelets. 4. Lymphoid stem cell can give rise to so-called "lymphoproliferative" malignancies such as acute lymphocytic leukemia 5. Myeloid Stem cell can give rise to so-called "myeloproliferative” malignancies such as acute myeloid leukemia II. Progenitor Cells:- CFU's(Colony Forming Units): 1. Derived from Multipotential StemCells.
    [Show full text]
  • GG Pur Bone Marrow Fm/C1 FNL-Br
    $50. 00 Bone Marrow Evaluation in Dogs and Cats Maxey L. Wellman, DVM, PhD, DACVP M. Judith Radin, DVM, PhD, DACVP Clinical Handbook Series Published by The Gloyd Group, Inc. Wilmington, Delaware © 2004 by Nestlé Purina PetCare Company. All rights reserved. Printed in the United States of America. Nestlé Purina PetCare Company: Checkerboard Square, Saint Louis, Missouri, 63188 First printing, 1999. This book is protected by copyright. ISBN 0-9678005-0-1 Table of Contents Introduction ......................................1 Part I Chapter 1 Normal Hematopoiesis ......................................5 Chapter 2 Indications for Bone Marrow Evaluation ................................13 Chapter 3 Procedures for Bone Marrow Aspiration and Biopsy ........................................................15 Chapter 4 Cytologic and Histologic Evaluation of the Bone Marrow Bone Marrow ....................................................17 Evaluation Part II in Dogs and Cats Chapter 5 Evaluation of Abnormal Bone Marrow ..........29 Maxey L. Wellman, DVM, PhD, DACVP Chapter 6 M. Judith Radin, DVM, PhD, DACVP Hematopoietic Neoplasia .................................43 Chapter 7 Case Studies .....................................................61 Part III Hematology Reference Ranges of the Dog and Cat .....................................................89 Index of Figures ...............................................90 Glossary of Terms ............................................96 Suggested Reading .........................................100 Clinical
    [Show full text]
  • Cleavage Fragments of the Third Complement Component (C3
    Leukemia (2007) 21, 973–982 & 2007 Nature Publishing Group All rights reserved 0887-6924/07 $30.00 www.nature.com/leu ORIGINAL ARTICLE Cleavage fragments of the third complement component (C3) enhance stromal derived factor-1 (SDF-1)-mediated platelet production during reactive postbleeding thrombocytosis M Wysoczynski1, M Kucia1, J Ratajczak1 and MZ Ratajczak1,2 1Stem Cell Biology Program, James Graham Brown Cancer Center, University of Louisville, Louisville, KY, USA and 2Department of Physiopathology, Pomeranian Medical University, Szczecin, Poland We hypothesized that the third complement component (C3) The a-chemokine stromal-derived factor-1 (SDF-1) has been cleavage fragments (C3a and des-ArgC3a) are involved in stress/ proposed as a new regulator of Megs maturation and platelet inflammation-related thrombocytosis, and investigated their 11–16 potential role in reactive thrombocytosis induced by bleeding. formation. A recently published study clearly demonstrated We found that platelet counts are lower in C3-deficient mice in that SDF-1, together with fibroblast growth factor-4 (FGF-4), À/À À/À response to excessive bleeding as compared to normal restored thrombopoiesis in Tpo and Mpl mice by directing littermates and that C3a and des-ArgC3a enhance stromal- trans-localization of megakaryocytic progenitors positive for derived factor-1 (SDF-1)-dependent megakaryocyte (Megs) CXCR4, the receptor for SDF-1, from the endostial to the migration, adhesion and platelet shedding. At the molecular vascular niche, thereby promoting survival, maturation to level, C3a stimulates in Megs MAPKp42/44 phosphorylation, 17 and enhances incorporation of CXCR4 into membrane lipid megakaryocytes and platelet production. rafts increasing the responsiveness of Megs to SDF-1.
    [Show full text]
  • Haematopoesis Solarsystem
    Haematopoiesis Peripheral blood Lymph nodes Bone marrow Haematopoietic stem cell B lymphocyte Pro-B Lymphoid-primed multipotent progenitor Common Multipotent lymphoid progenitor progenitor Lymphoblast Monoblast Promonocyte Granulocyte- monocyte Monocyte Common myeloid progenitor progenitor Bone marrow NK-cell Neutrophilic Neutrophilic myelocyte metamyelocyte Neutrophilic Pro-T Myeloblast band cell Neutrophilic granulocyte Promyelocyte Thymus Eosinophilic myelocyte Megakaryocyte-erythroid T lymphocyte progenitor Bone marrow Proerythroblast Eosinophilic metamyelocyte Megakaryoblast Basophilic Eosinophilic myelocyte band cell Basophilic erythroblast Peripheral blood Eosinophilic granulocyte Basophilic metamyelocyte Polychromatic Promegakaryocyte erythroblast Basophilic Orthochromatic band cell erythroblast The illustration of haematopoiesis as a ‘solar system’ model is based on the idea of a high plasticity of haematopoiesis and an intimate relationship Peripheral blood between various types of progenitor cells.* Many progenitors share common transcription factors. The proximity of radial sectors of different cell Megakaryocyte lineages depicts how close the ontogeny of these lineages is. Gradual colour change reflects the potential of early and intermediate progenitors Basophilic to follow different cells’ fates. granulocyte Reticulocyte Reticulated platelet Haematopoietic stem cell Common lymphoid progenitor Granulocyte-monocyte progenitor A true stem cell that has a potential to self-renew The early progenitor committed to lymphoid This intermediate progenitor is committed and differentiate into any lineage of blood cells. lineage. Gives rise to precursors of T and B to monocytic and granulocytic lineages. lymphocytes and natural killer cells. Lymphoid Multipotent progenitor progenitors leave the bone marrow for matu- Megakaryocyte-erythroid progenitor Can give rise to any blood cell lineage. ration in the thymus and lymph nodes. Intermediate progenitor that can only differ- entiate into a red blood cell or megakaryocyte.
    [Show full text]