DOCSLIB.ORG

Mobilized Peripheral Blood Stem Cells Provide Rapid Reconstitution but Impaired Long-Term Engraftment in a Mouse Model

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mobilized Peripheral Blood Stem Cells Provide Rapid Reconstitution but Impaired Long-Term Engraftment in a Mouse Model Bone Marrow Transplantation (2007) 39, 401–409 & 2007 Nature Publishing Group All rights reserved 0268-3369/07 $30.00 www.nature.com/bmt ORIGINAL ARTICLE Mobilized peripheral blood stem cells provide rapid reconstitution but impaired long-term engraftment in a mouse model JSG Yeoh, A Ausema, P Wierenga, G de Haan and R van Os Department of Cell Biology, Section Stem Cell Biology, University Medical Centre Groningen, Groningen, The Netherlands In this study, we use competitive repopulation to compare become the primary source of hematopoietic stem cells for the quality and frequency of stem cells isolated from clinical transplantation over the past 10 years.4 mobilized blood with stem cells isolated from bone marrow Although multiple hematopoietic growth factors are (BM) in a mouse model. LinÀSca-1 þ c-Kit þ (LSK) cells capable of inducing mobilization of hematopoietic pro- were harvested from control BM and peripheral blood of genitors, granulocyte colony-stimulating factor (G-CSF) is mice following granulocyte colony-stimulating factor (G- at present the most used mobilizing molecule in clinical CSF) administration. LSK cells were used because oftheir protocols.5 The ability of G-CSF to mobilize BM resemblance to human CD34 þ cells. We confirmed that precursors and long-term repopulating cells was initially transplantation ofphenotypically defined mobilized per- shown in preclinical studies. Molineux et al.6 observed a ipheral blood (MPB) stem cells results in rapid recovery of marked increase in the colony-forming unit spleen pool in blood counts. However, in vitro results indicated that LSK the peripheral blood of mice treated with repeated doses of cells purified from MPB had lower cobblestone area- G-CSF. In addition, it was observed that G-CSF alone and forming cell day 35 activity compared to BM. Addition- in combination with stem cell factor or interleukin-7 ally, evaluation ofchimerism after co-transplantation of mobilizes hematopoietic precursors capable of both radio- LSK cells purified from blood and BM revealed that MPB protection and generating sustained lymphohematopoiesis stem cells contained 25-fold less repopulation potential in transplanted recipients.7 In vitro data from human compared to BM stem cells. Competitive repopulating patients appear consistent with the concept that the quality unit frequency analysis showed that freshly isolated MPB of human mobilized peripheral blood (MPB) progenitor LSK cells have 8.8-fold fewer cells with long-term cells is at least equivalent to that corresponding to BM repopulating ability compared to BM LSK cells. Second- grafts.8,9 ary transplantation showed no further decline in contribu- Surprisingly, despite the prevalent use of hematopoietic tion ofhematopoiesis relative to BM. We conclude that stem cell mobilization in clinical transplantation, few the reduced frequency of stem cells within the LSK reports exist describing the competitive repopulating population ofMPB, rather than poorer quality, causes the quality of mobilized stem cells compared to BM stem cells reduced repopulation potential. following G-CSF treatment. Most available reports only Bone Marrow Transplantation (2007) 39, 401–409. outlined the differences in the kinetics and efficiency of doi:10.1038/sj.bmt.1705601; published online 12 February 2007 engraftment, in homing properties and in cell cycle profiles Keywords: granulocyte colony-stimulating factor; mobi- between mobilized blood stem cells and those isolated from lized peripheral blood stem cells; bone marrow stem cells; resting BM.10–13 competitive repopulating unit; long-term engraftment In view of the increased use of peripheral blood stem cells in clinical transplant settings, it is of relevance to investigate long-term functioning of stem cells isolated from different sources. In this study, we directly assessed the function of MPB stem cells compared to control BM Introduction stem cells when co-transplanted in a single recipient mouse in an in vivo competitive repopulation assay. We show a The initial source of hematopoietic cells used for trans- 1 reduced frequency of repopulating stem cells in purified plantation was bone marrow (BM). However, owing to the G-CSF-MPB LinÀSca-1 þ c-Kit þ (LSK) stem cells, which faster regeneration of both circulating neutrophils (9–11 2,3 caused a 25-fold reduction in repopulation potential days) and platelets, peripheral blood stem cells have compared with BM LSK cells. Correspondence: Dr R van Os, Department of Cell Biology, Section Materials and methods Stem Cell Biology, University Medical Centre Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands. E-mail: [email protected] Mice Received 14 September 2006; revised 29 November 2006; accepted 5 Female C57BL/6 (B6), C57BL/6.SJL (CD45.1), (C57BL/ January 2007; published online 12 February 2007 6 Â C57BL6.SJL) F1 (CD45.1/2) or C57BL/6-Tg(ACTB- MPB stem cells have impaired long-term engraftment JSG Yeoh et al 402 EGFP)10sb/J transgenic GFP (GFP) mice were used as national IBL 637 137Cs-source, 20–24 h before transplant donors, competitors or recipients of blood and marrow ation. For competitive repopulation determination, stem cells depending on the experimental model. CD45.1 unfractionated cells or MPB LSK cells, were mixed with and transgenic GFP mice were originally obtained from the competitor cells (unfractionated or LSK BM cells) and Jackson Laboratory (Bar Harbor, ME, USA) and bred in intravenously transplanted into recipient mice. Each our local animal facility. Wild-type female B6 mice were transplant group consisted of 8–10 recipients. Following purchased from Harlan (Horst, The Netherlands) and transplantation, blood samples (60 ml) were taken monthly maintained under clean conventional conditions in the to determine donor chimerism. Levels of chimerism animal facilities of University Medical Centre Groningen were determined by detecting the presence of GFP þ or (The Netherlands). Mice were fed ad libitum with food CD45.1 þ and CD45.2 þ cells in transplanted mice. To pellets and acidified tap water (pH ¼ 2.8). All animal detect CD45.1 þ and CD45.2 þ cells, cells were stained with procedures were approved by the local animal ethics anti-CD45.2 (FITC) and anti-CD45.1 (PE) antibodies (BD committee of the University Medical Centre Groningen. Pharmingen, San Diego, CA, USA) for 30 min and analyzed on a flow cytometer (fluorescence-activated cell Mobilization and harvesting of stem cells sorter (FACS) Calibur; Becton Dickinson Biosciences, San BM cells were harvested by flushing the femoral shaft with Jose, CA, USA). In addition, the competitive repopulating a-Modified Eagle’s Medium (GibcoBRL, Invitrogen, CA, index (CRI) was determined. CRI is a relative measure of USA) supplemented with 2% fetal calf serum (FCS; the competitive ability of test cells to that of fresh BM cells. GibcoBRL, Invitrogen, CA, USA). Pegylated G-CSF The CRI was calculated by taking the ratio of white blood (Neulasta) (Amgen, Thousand Oaks, CA, USA) (250 mg/ cell (WBC) derived from mobilized blood cells to BM cells kg/mouse) was used to mobilize stem cells.14 Two doses of in the circulation and dividing it by the ratio of mobilized G-CSF were subcutaneously administered to donor mice at blood cells to BM cells transplanted. A CRI value of one day À6 and day À3 before stem cell harvest. MPB cells indicates by definition that MPB cells and BM cells have were harvested by cardiac puncture. Approximately 1 ml of equal competitive ability. blood was collected and was diluted with 4 ml of Iscove’s modified Dulbecco’s modified Eagle’s medium (IMDM; CRU assay GibcoBRL, Paisley, Scotland) supplemented with 5% FCS B6-recipient mice were transplanted with a series of diluted (GibcoBRL, Invitrogen, CA, USA) and heparin (25 IU) CD45.1 LSK cells (1200, 600 and 300) from mobilized (Leo Pharma, Breda, Netherlands). The collected blood cell blood and control BM and with a fixed number of B6 suspension (5 ml) was centrifuged over an equal volume of competitor cells (5 Â 105). Twelve weeks after transplant- Lympholyte-M (Cedarlane Laboratories Ltd, Hornby, ation, donor cell contribution in the peripheral blood was Canada) at 400 Â g for 30 min at room temperature. After determined. Recipients with a contribution of X5% in both centrifugation, the mononuclear cells within the opaque myeloid and lymphoid lineages were considered to be interface layer were isolated and washed in IMDM/5% positive. FCS for 5 min at 2000 r.p.m. at 41C. Alternatively, red To evaluate and quantify the repopulating potential of blood cells were lysed using ammonium chloride (NH4Cl) mobilized blood LSK cells and control BM LSK cells, the without prior density separation. Nucleated cells were frequency of competitive repopulation units (CRU) was measured on a Coulter Counter Model Z2 (Coulter calculated. CRU frequencies per 1000 LSK were calculated Electronics, Hialeah, FL, USA). from the resultant percentage positive recipients by limiting dilution analysis procedures which uses Poisson statistics.16 Isolation of LSK cells BM and MPB cells were stained as described previously15 CAFC assays with biotinylated lineage-specific Mouse Lineage Panel CAFC assays were performed as described17–19 to assess the antibodies, containing anti-CD45R, anti-CD11b, anti- number of hematopoietic progenitor cells (CAFC day 7) or TER119, anti-Gr-1 and anti-CD3e (BD Pharmingen, San more primitive stem cells (CAFC day 35) in MPB Diego, CA, USA), fluorescein isothiocyanate (FITC)-anti- stem cells. Sca-1 and APC-anti-c-kit (BD Pharmingen, San Diego, CA, USA). Biotinylated antibodies were visualized with Secondary transplantations streptavidin–phycoerythrin (PE) (Pharmingen, San Diego, In one of the competitive repopulation experiments, in CA, USA). After antibody staining, cells were sorted by a which recipients were transplanted in different ratios with MoFlow cell sorter (DakoCytomation, Fort Collins, CO, CD45.1 MPB LSK cells and CD45.2 BM LSK competitor À þ þ USA). LSK and Lin non-Sca-1 c-Kit cells were sorted cells, mice were killed for secondary transplantation.
Load more

Recommended publications
  • Peripheral Blood Cells Changes After Two Groups of Splenectomy And
    ns erte ion p : O y p H e f Yunfu et al., J Hypertens (Los Angel) 2018, 7:1 n o l A a DOI: 10.4172/2167-1095.1000249 c c n r e u s o s J Journal of Hypertension: Open Access ISSN: 2167-1095 Research Article Open Access Peripheral Blood Cells Changes After Two Groups of Splenectomy and Prevention and Treatment of Postoperative Complication Yunfu Lv1*, Xiaoguang Gong1, XiaoYu Han1, Jie Deng1 and Yejuan Li2 1Department of General Surgery, Hainan Provincial People's Hospital, Haikou, China 2Department of Reproductive, Maternal and Child Care of Hainan Province, Haikou, China *Corresponding author: Yunfu Lv, Department of General Surgery, Hainan Provincial People's Hospital, Haikou 570311, China, Tel: 86-898-66528115; E-mail: [email protected] Received Date: January 31, 2018; Accepted Date: March 12, 2018; Published Date: March 15, 2018 Copyright: © 2018 Lv Y, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Objective: This study aimed to investigate the changes in peripheral blood cells after two groups of splenectomy in patients with traumatic rupture of the spleen and portal hypertension group, as well as causes and prevention and treatment of splenectomy related portal vein thrombosis. Methods: Clinical data from 109 patients with traumatic rupture of the spleen who underwent splenectomy in our hospital from January 2001 to August 2015 were retrospectively analyzed, and compared with those from 240 patients with splenomegaly due to cirrhotic portal hypertension who underwent splenectomy over the same period.
    [Show full text]
  • Combined Effect of Granulocyte-Colony-Stimulating
    J Korean Med. 2016;37(4):36-44 http://dx.doi.org/10.13048/jkm.16046 pISSN 1010-0695 • eISSN 2288-3339 Original Article Combined Effect of Granulocyte-Colony-Stimulating Factor-Induced Bone Marrow-Derived Stem Cells and Red Ginseng in Patients with Decompensated Liver Cirrhosis (Combined Effect of G-CSF and Red Ginseng in Liver Cirrhosis) Hyun Hee Kim1, Seung Mo Kim2, Kyung Soon Kim2, Min A Kwak2, Sang Gyung Kim3, Byung Seok Kim1, Chang Hyeong Lee1 1Department of Internal Medicine, Catholic University of Daegu School of Medicine 2Department of Internal Medicine, College of Korean Medicine, Daegu Haany University 3Department of Laboratory Medicine, Catholic University of Daegu School of Medicine Objectives: Granulocyte-colony-stimulating factor (G-CSF) mobilized bone marrow (BM)-derived hematopoietic stem cells could contribute to improvement of liver function. In addition, liver fibrosis can reportedly be prevented by the Rg 1 component of red ginseng. This study investigated the combined effect of G-CSF and red ginseng on decompensated liver cirrhosis. Methods: Four patients with decompensated liver cirrhosis were injected with G-CSF to proliferate BM stem cells for 4 days (5 μg/kg bid subcutaneously) and followed-up for 3 months. The patients also received red ginseng for 4 days (2 tablets tid per os). We analyzed Child-Pugh scores, Model for End-Stage Liver Disease (MELD) scores and cirrhotic complications. Results: All patients showed marked increases in White blood cell (WBC) and CD34+ cells in the peripheral blood, with a peak time of 4 days after G-CSF injection. Spleen size also increased after G-CSF injection, but not severely.
    [Show full text]
  • Factors Influencing Platelet Recovery After Blood Cell Transplantation in Multiple Myeloma
    Bone Marrow Transplantation, (1997) 20, 375–380 1997 Stockton Press All rights reserved 0268–3369/97 $12.00 Factors influencing platelet recovery after blood cell transplantation in multiple myeloma MA Gertz1, MQ Lacy1, DJ Inwards1, AA Pineda2, MG Chen3, DA Gastineau1, A Tefferi1, RA Kyle1 and MR Litzow1 1Division of Hematology and Internal Medicine, 2Division of Transfusion Medicine, and 3Division of Radiation Oncology, Mayo Clinic and Mayo Foundation, Rochester, MN, USA Summary: cells result in durable engraftment after transplantation.2 Hematopoietic recovery can be accelerated by the use of We sought to determine factors that impact on the blood stem cells mobilized with growth factors or chemo- recovery of platelets after blood cell transplantation in therapy (or both). Hematopoietic recovery is more rapid patients with multiple myeloma. We performed retro- after mobilized peripheral blood stem cell transplantation spective analyses in 51 patients undergoing blood cell than after autologous bone marrow transplantation, and transplantation for multiple myeloma. The pro- hematopoietic recovery has been shown to be sustained, portional-hazards model was applied to determine sig- indicating that accessory cells found in the bone marrow nificant risk factors. Of 51 transplants, 14 patients failed are not required for durable engraftment.3 The more rapid to achieve a platelet count of 50 × 109/l. Median time to engraftment with mobilized stem cells is likely due to an a neutrophil count of 0.5 × 109/l was 10.5 days. Median increase in committed
    [Show full text]
  • Blood Lab First Week
    Blood Lab First Week This is a self propelled powerpoint study atlas of blood cells encountered in examination of the peripheral blood smear. It is a requirement of this course that you begin review of these slides with the first day of class in order to familiarize yourself with terms used in describing peripheral blood morphology. Laboratory final exam questions are derived directly from these slides. The content of these slides may duplicate slides (1-37) listed on the Hematopathology Website. You must familiarize yourself with both sets of slides in order to have the best learning opportunity. Before Beginning This Slide Review • Please read the Laboratory information PDF under Laboratory Resources file to learn about – Preparation of a blood smear – How to select an area of the blood smear for review of morphology and how to perform a white blood cell differential – Platelet estimation – RBC morphology descriptors • Anisocytosis • Poikilocytosis • Hypochromia • Polychromatophilia Normal blood smear. Red blood cells display normal orange pink cytoplasm with central pallor 1/3-1/2 the diameter of the red cell. There is mild variation in size (anisocytosis) but no real variation in shape (poikilocytosis). To the left is a lymphocyte. To the right is a typical neutrophil with the usual 3 segmentations of the nucleus. Med. Utah pathology Normal blood: thin area Ref 2 Normal peripheral blood smear. This field is good for exam of cell morphology, although there are a few minor areas of overlap of red cells. Note that most cells are well dispersed and the normal red blood cell central pallor is noted throughout the smear.
    [Show full text]
  • Causes of Peripheral Blood Cytopenias in Patients with Liver Cirrhosis Portal Hypertension and Clinical Significances
    Open Journal of Endocrine and Metabolic Diseases, 2014, 4, 85-89 Published Online April 2014 in SciRes. http://www.scirp.org/journal/ojemd http://dx.doi.org/10.4236/ojemd.2014.44010 Causes of Peripheral Blood Cytopenias in Patients with Liver Cirrhosis Portal Hypertension and Clinical Significances Yunfu Lv Department of General Surgery, People’s Hospital of Hainan Province, Haikou, China Email: [email protected] Received 18 March 2014; revised 12 April 2014; accepted 19 April 2014 Copyright © 2014 by author and Scientific Research Publishing Inc. This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/ Abstract Liver cirrhosis portal hypertension patients to reduce the number of blood cells are common in clinical, and often affect the prognosis. This paper discusses cirrhotic portal hypertension patients complicated by the reason of the decrease in the number of peripheral blood cells and what is the clinical significance of these reasons so as to provide theoretical support for the choice of treat- ment. Splenomegaly and hypersplenism caused should be the main reason for reducing the num- ber of blood cells, but not all, other reasons are alcohol and virus inhibition of bone marrow, liver function impairment, autoimmune damage and loss of blood, etc. If it is a function of the spleen hyperfunction caused by blood cells decreases, blood should rise to normal after splenectomy, or consider other reason or there are other reasons at the same time. Keywords Liver Cirrhosis Portal Hypertension, Peripheral Blood Cytopenias, Causes, Cilinical Significances 1. Introduction There are approximately 350 million carriers of hepatitis B virus (HBV) worldwide, and more than half of them are in the Asia-Pacific region.
    [Show full text]
  • Identifying Abnormalities in Peripheral Blood Cells
    EDUCATIONAL COMMENTARY – IDENTIFYING ABNORMALITIES IN PERIPHERAL BLOOD CELLS Educational commentary is provided through our affiliation with the American Society for Clinical Pathology (ASCP). To obtain FREE CME/CMLE credits click on Earn CE Credits under Continuing Education on the left side of the screen. Learning Outcomes Upon completion of this exercise, participants will be able to: describe the morphologic features of normal peripheral blood leukocytes. identify abnormalities in red blood cells (RBCs) that are characteristic of hereditary spherocytosis. discuss the pathophysiology of hereditary spherocytosis. An 8 year old female was seen in the pediatric hematology clinic. Laboratory findings included: WBC= 13.6 x 109/L, RBC= 2.63 x 1012/L, Hgb= 8.5 g/dL, Hct= 22%, MCV= 85.0 fL, MCH= 32.4 pg, MCHC= 38.1 g/dL, RDW= 21.4%, and Platelets= 897 x 109/L. This patient has been diagnosed with hereditary spherocytosis. Normal leukocytes and abnormalities in erythrocytes that may be seen in this condition are presented in the images. BCI-01 shows an eosinophil. Eosinophils characteristically have large, red-orange cytoplasmic granules. These granules are numerous and uniform in size. The nucleus of an eosinophil is often bilobed, although in this image, it is round. The chromatin is generally dense and clumped. American Proficiency Institute – 2011 1st Test Event EDUCATIONAL COMMENTARY – IDENTIFYING ABNORMALITIES IN PERIPHERAL BLOOD CELLS (cont.) The cell identified in BCI-02 is a segmented neutrophil. Neutrophils usually have 2 to 5 nuclear lobes connected by thin filaments of chromatin. The chromatin is coarse and densely clumped. In contrast to the eosinophil shown in BCI-01, segmented neutrophils have numerous small granules that normally stain light-violet or pink.
    [Show full text]
  • Peripheral Blood Cell Variations in Cirrhotic Portal Hypertension Patients with Hypersplenism Yun-Fu Lu1, Xin-Qiu Li2*, Xiao-Yu Han1, Xiao-Guang Gong1, Shun-Wu Chang1
    Asian Pacific Journal of Tropical Medicine (2013)663-666 663 Contents lists available at ScienceDirect Asian Pacific Journal of Tropical Medicine journal homepage:www.elsevier.com/locate/apjtm Document heading doi: Peripheral blood cell variations in cirrhotic portal hypertension patients with hypersplenism Yun-Fu Lu1, Xin-Qiu Li2*, Xiao-Yu Han1, Xiao-Guang Gong1, Shun-Wu Chang1 1Department of Surgery, People s Hospital of Hainan Province, Haikou 570311, China 2Department of Thyroid, Mammary' Gland and Blood Vessel surgery, Renmin Hospital, Hubei University of Medicine, Shiyan 442000, China ARTICLE INFO ABSTRACT Article history: Objective: To explore peripheral blood cell variations in hepatic cirrhosis portal hypertension Methods: Received 10 May 2013 patients with hypersplenism. Clinical data of 322 hypersplenism patients with Received in revised form 15 June 2013 decreased peripheral blood cells, admitted with cirrhotic portal hypertension, was retrospectively Accepted 15 July 2013 Results: 17 I 64% (206 322) 2 Available online 20 August 2013 studied over the last years. n / of patients, more than kinds of blood cell were decreased, including 89 cases of pancytopenia (43.2%), 52 cases of WBC + PLT decrease (25.2%), 29 cases of RBC + PLT decrease (14.1%), and 36 cases of WBC + RBC decrease (17.5%); in 36% (116/322) of patients, single type blood cell decrease occurred, including 31 cases of PLT Keywords: decrease (26.7%), 29 cases of WBC decrease (25%) and 56 cases of RBC decrease (48.3%). Of 227 Hepatic cirrhosis portal hypertension routine bone marrow examinations, bone marrow hyperplasia was observed in 118 cases (52.0%), Peripheral blood cell the remainder showed no hyperplasia.
    [Show full text]
  • The Acute Phase Reaction and Its Prognostic Impact In
    biomedicines Article The Acute Phase Reaction and Its Prognostic Impact in patients with Head and Neck Squamous Cell Carcinoma: Single Biomarkers Including C-Reactive Protein Versus Biomarker Profiles Helene Hersvik Aarstad 1,2, Svein Erik Emblem Moe 2, Øystein Bruserud 1,3, Stein Lybak 2, 2,4, , 3, Hans Jørgen Aarstad * y and Tor Henrik Anderson Tvedt y 1 Department of Clinical Science, Faculty of Medicine, University of Bergen, 5021 Bergen, Norway; [email protected] (H.H.A.); [email protected] (Ø.B.) 2 Department of Otolaryngology/Head and Neck Surgery, Haukeland University Hospital, 5021 Bergen, Norway; [email protected] (S.E.E.M.); [email protected] (S.L.) 3 Section for Hematology, Department of Medicine, Haukeland University Hospital, 5021 Bergen, Norway; [email protected] 4 Department of Clinical Medicine, Faculty of Medicine, University of Bergen, 5021 Bergen, Norway * Correspondence: [email protected]; Tel.: +47-5597-2664 These two authors shared the last authorship equally. y Received: 6 September 2020; Accepted: 13 October 2020; Published: 14 October 2020 Abstract: C-reactive protein (CRP) has a prognostic impact in head and neck squamous cell carcinoma (HNSCC). However, the acute phase reaction involves many other proteins depending on its inducing events, including various cytokines that can function as reaction inducers. In the present study, we compared the pretreatment acute phase cytokine profile for 144 patients with potentially curative HNSCC. We investigated the systemic levels of interleukin (IL)6 family mediators (glycoprotein (gp130), IL6 receptor (R)α, IL6, IL27, IL31, oncostatin M (OSM), ciliary neurotrophic factor (CNTF)), IL1 subfamily members (IL1R antagonist (A), IL33Rα), and tumor necrosis factor (TNF)α.
    [Show full text]
  • Platelet Counting: Ugly Traps and Good Advice
    Journal of Clinical Medicine Review Platelet Counting: Ugly Traps and Good Advice. Proposals from the French-Speaking Cellular Hematology Group (GFHC) Véronique Baccini 1,* , Franck Geneviève 2, Hugues Jacqmin 3, Bernard Chatelain 3, Sandrine Girard 4, Soraya Wuilleme 5, Aurélie Vedrenne 6, Eric Guiheneuf 7 , Marie Toussaint-Hacquard 8, Fanny Everaere 9, Michel Soulard 10, Jean-François Lesesve 8 and Valérie Bardet 11 1 Laboratoire d’hématologie, CHU de la Guadeloupe, INSERM UMR S_1134, 97159 Pointe-à-Pitre, France 2 Fédération Hospitalo-Universitaire ‘Grand Ouest Against Leukemia’ (FHU GOAL), 49033 Angers, France; [email protected] 3 Université Catholique de Louvain, CHU UCL Namur, Laboratoire d’hématologie, Namur Thrombosis and Hemostasis Center, 5530 Yvoir, Belgium; [email protected] (H.J.); [email protected] (B.C.) 4 Hospices Civils de Lyon, Centre de biologie et pathologie Est, Service d’hématologie biologique, 69500 Bron, France; [email protected] 5 Laboratoire d’Hématologie, Institut de Biologie, CHU de Nantes; 44093 Nantes CEDEX, France; [email protected] 6 Service de biologie clinique, Hôpital Foch, 92150 Suresnes, France; [email protected] 7 Service d’Hématologie Biologique, CHU Amiens-Picardie, 80054 Amiens CEDEX, France; [email protected] 8 Hématologie Biologique, CHRU Nancy, 54511 Vandoeuvre, France; [email protected] (M.T.-H.); [email protected] (J.-F.L.) 9 Audolys Biologie, 62219 Longuenesse, France; [email protected] 10 Plateau
    [Show full text]
  • Toxicologic Pathology – Immune System of Laboratory Animals
    Toxicologic Pathology – Immune System of Laboratory Animals Klaus Weber, PhD, DVM, MSBiol AnaPath GmbH, Switzerland In Cooperation with BSL Scientific Laboratories GmbH, Planegg, Germany Immune System: What it is? Compartimentation Guidelines? • Detailed strategy • All lymphoid tissues to be examined (incl. Peyer’s patches) • Immunohistochemistry superior to Facscan • Interpretation of stress-related effects are necessary Compartimentation Parameter Specific Component Hematology Total and absolute differential leukocyte counts Clinical Globulin levels1 and A/G ratios Chemistry Gross Pathology Lymphoid organs / tissues Organ Weights Thymus, spleen (optional: lymph nodes) Histology Thymus, spleen, draining lymph node and at least one additional lymph node, bone marrow2, Peyer’s patch3, BALT4, NALT4 1Unexplained alterations in globulin levels could call for measurements of immunoglobulins 2Unexplained alterations in peripheral blood cell lines or histopathological findings might suggest that cytologic evaluation of the bone marrow would be appropriate 3Oral administration only 4For inhalation or nasal route only Lymph Nodes Functional Structure • Lymphocytes of the whole body turns over 10 to 48x/24 hrs • DC - Dendritic cells as APC’s (loosing ability to bind antigens during travel to lymph nodes but gaining ability to present) – presenting to T-cells with subsequent proliferation after 1-2 days • FDC - Follicular dendritic cells: APC’s that present to B-cells • Germinal centers formed by B-cells where they are in contact with FDC’s after
    [Show full text]
  • Modification of Rhodamine Staining Allows Identification of Hematopoietic Stem Cells with Preferential Short-Term Or Long-Term Bone Marrow-Repopulating Ability J
    Proc. Natl. Acad. Sci. USA Vol. 92, pp. 8901-8905, September 1995 Medical Sciences Modification of rhodamine staining allows identification of hematopoietic stem cells with preferential short-term or long-term bone marrow-repopulating ability J. MARK J. M. ZIJLMANS*t, JAN W. M. VISSER*t, KARIN KLEIVERDA§, PHILIP M. KLUIN§, ROEL WILLEMZE*, AND WILLEM E. FIBBE* *Laboratory of Experimental Hematology, Department of Hematology, and §Department of Pathology, University Medical Center, 2300 RC Leiden, The Netherlands; and tLindsley F. Kimball Research Institute, Laboratory of Stem Cell Biology, New York Blood Center, New York, NY 10021 Communicated by J. J. van Rood, University Hospital, Leiden, The Netherlands, June 12, 1995 ABSTRACT We have developed a modified rhodamine In addition to methods exploiting surface marker expres- (]Rho) staining procedure to study uptake and efflux in sion, other techniques are based on functional differences. murine hematopoietic stem cells. Distinct populations of One of these involves the use of rhodamine 123 (Rho), a Rho++ (bright), Rho+ (dull), and Rho- (negative) cells could supravital fluorescent dye that accumulates intracellularly and be discriminated. Sorted Rho- cells were subjected to a that may bind to mitochondria (5, 6). Cells with smaller or less second Rho staining procedure with the P-glycoprotein block- active mitochondria exhibit reduced uptake of Rho (6). Rho ing agent verapamil (VP). Most cells became Rho positive efflux from the cell is mediated by the transmembrane carrier [Rho-/Rho(VP)+ cells] and some remained Rho negative protein P glycoprotein, encoded by the multidrug-resistance [Rho-/Rho(VP)- cells]. These cell fractions were character- gene (7-9).
    [Show full text]
  • Platelet Morphology Received January 20, 2020; Accepted March 28, 2020 Introduction
    J Lab Med 2020; 44(5): 231–239 Review Christoph Robier* Platelet morphology https://doi.org/10.1515/labmed-2020-0007 Received January 20, 2020; accepted March 28, 2020 Introduction Abstract The examination of a peripheral blood (PB) smear should be requested in every case of unexplained thrombocytope- Background: The examination of a peripheral blood smear nia or thrombocytosis. First, the morphological analysis is mandatory in case of unexplained thrombocytopenia should include the estimation of the number of platelets or thrombocytosis. First, the number of platelets should in order to confirm the platelet count determined by the be estimated in order to confirm the platelet count deter- haematology analyser, and to rule out causes of spuriously mined by the haematology analyser, and to rule out causes low platelet counts due to platelet aggregates, platelet sat- of spuriously low or elevated platelet counts. Second, the ellitism or platelet phagocytosis [1–4]. The identification size and morphological features of the platelets, which of pseudothrombocytopenia (PTP) due to anticoagulant- may provide information on the underlying cause of the induced platelet aggregation is crucial, because a misin- low or enhanced platelet count, have to be assessed. terpretation as a “true” thrombocytopenia may lead to Content: This review summarizes the physiological and serious diagnostic and therapeutic consequences such as pathological features of platelet size and morphology, bone marrow biopsy, initiation of corticosteroid therapy, circulating megakaryocytes, micromegakaryocytes and platelet transfusion or even splenectomy [2]. PTP can be megakaryoblasts, and provides an overview of current caused by various anticoagulants such as ethylenedi- guidelines on the reporting of platelet morphology.
    [Show full text]