HAEMATOPOIESIS

Dr Rosline Hassan Hematology Department School of Medical Sciences Universiti Sains Malaysia

Organization of the hematopoietic system

Haematopoiesis

Definition : production of red blood cells, white cells and platelets 175 billion red blood cells/day 70 billion granulocytes/day (neutrophils, eosinophils, basophils) 175 billion platelets/day Capable of 5-10 fold increase in production

Embryogenesis
Dev. Foetus age Sites 0-2 months Yolk sac

2-7 months Liver & spleen

Infants
Adults

Bone marrow (all bones)

Mesoblastic phase

2nd week - 3rd month Cell clusters in yolk sac 2nd - 6th month Mainly liver with supplementation from spleen 6th month adult Red bone marrow

Hepatic phase

Myeloid phase

Anatomy of Marrow

Volume of bone marrow at birth: 1.5% of body weight Volume of bone marrow as an adult: 4.5% of body weight

Haematopoiesis

Anatomy of Marrow
Histology Vascular Compartment Hematopoietic Compartment

hematopoietic stem cells Precursor cells growth factor hemotopoietic microenvironment (stroma) site for formation and maturation

Cellularity 30-70%

Haematopoietic cells

Stem Cells

Pluripotential Stem Cells

Multipotential Stem Cells ( Lymphoid, Myeloid Stem Cells) Progenitor Cells - CFU's (Colony Forming Units)

Stem cell

Multipotential stem cell

Progenitor cells

Haematopoietic cells

Precursor Cells - Blasts and their progeny

First morphologically identifiable cells

Little if any self-renewal

Haematopoietic cells

Mature Effector Cells

Red Cells carry oxygen, carbon dioxide; lifespan 120 days Neutrophils phagocytosis, killing Monocytes phagocytosis, killing, antigen presentation Lymphocytes immune response Platelets haemostasis

Normal Peripheral Blood

Monocyte Neutrophil

Lymphocyte

Basophil

Microenvironment
Forms a suitable environment for stem cell growth and development. Composed of stromal cells & a microvascular network

Bone Marrow Stromal Cells - key role in marrow Fibroblasts, fat cells, endothelial cells, reticulum cells and macrophage Express Adhesion Molecules Secrete Growth Factors and extracellular molecules (collagen, glycoproteins & glycosaminoglycans) forming extracellular matrix Stromal matrix Physical Support for Hematopoietic Cells

Haemopoiesis occur within microenvironment Stem cells are in contact with other cell (e.g macrophage, fibroblast) Cell-cell communication is by binding via cell surface receptors, to adhesion molecules & secretion of cytokines & GFs These binding lead to proliferation, differentiation & apoptosis

vessel

Fat cells

bone

Hemopoietic cells

Haematopoietic Growth Factors

Glycoprotein hormones Function : Progenitor cells

e.g : IL-1, TNF, IL-3, erythropoietin

Mature blood cells

Proliferation Differentiation Maturation

Functional activation Prevention of apoptosis

Act locally or circulate in plasma Produced by :

lymphocytes monocytes macrophages

endothelial cells fibroblasts

Classification

Non lineage specific GF

Act on pluripotential & multipotential stem cells to initiate self renewal & differentiation e.g :IL-3, GM-CSF

Lineage specific GF

Act on progenitor cells, involve in differentiation & maturation of cells e.g :CSF, M-CSF, erythropoetin & thrombopoietin
Release by lymphocytes & monocytes e.g : interleukin-1 (IL-1)

Lymphokine and monokines

Haematopoiesis

Erythropoiesis

Occurs near the sinus apertures Seen as erythroblastic islands surround the macrophages Matured red cell squeeze through the aperture leaving their nuclei behind
Occurs deep in the cords Mature cells become motile at the metamyelocyte stage

Granulopoiesis

Lymphopoiesis

Leave the bone marrow and complete differentiation in the lymph nodes (B cells) and thymus (T cells)
Occurs along the outside surface of the vascular sinuses Release strings of cytoplasmic platelet directly into the circulation.

Thrombopoiesis

Erythropoiesis

RBC development takes about 7 days Involves 3 to 4 mitotic cell divisions Each stem cell gives rise to 8 or 16 cells Stages of RBC development characterized by 1. gradual appearance of haemoglobin 2. disappearance of ribonucleic acid (RNA) 3. progressive degeneration of nucleus 4. gradual loss of cytoplasmic organelles e.g mitochondria 5. gradual reduction in cell size

Substances required for erythropoiesis : i. Metal (e.g: iron, mangenese & cobalt) ii. Vitamin (e.g: B12, Folate, Vit C, Vit E, Vit B6 (pyridoxine), thiamine, riboflavin & pantothenic acid) iii. Amino acid iv. Hormones (e.g: stem cell factor, IL-3, GMCSF, erythropoietin, androgen & thyroxine)

Life cycle of RBCs

2.

3.

4.

5.

6.

Kidneys respond to lower oxygen concentration in blood releasing erythropoietin (EPO). EPO travels to red marrow and stimulates production of RBCs Red marrow produces RBCs from stem cells RBCs squeeze through vessel membrane to enter circulation Heart and lungs work to supply oxygenation of RBCs. Damaged or old RBCs destroyed by the spleen.

Leukopoiesis

Leukopoiesis (white blood cell formation) encompasses both granulopoiesis and agranulopoiesis. Granulopoiesis occurs in the bone marrow of healthy adults. Kinetics of Neutrophil Production - 14 days from myeloblast to neutrophil.

Granulopoiesis

3 types of granulocytes (derive from a single precursor CFU-G)

Neutrophils Basophils Eosinophils

The structural changes include

4.

Decrease in cell size Condensation of nuclear chromatin Changes in nuclear shape (flattening indentation lobulation Accumulation of cytoplasmic granules

Stages of maturation : 1. Myeloblast 2. Promyelocyte (10 granules) 3. Myelocyte (20 granules)

Last mitotically active stage in granulopoiesis.

4. 5.

Metamyelocyte Band neutrophil / stab form

Lymphopoiesis

Production of lymphocytes. Lymphoid stem cells give rise to the major functional classes of lymphocytes : i. B cells (Undergo differentiation to become plasma cells and produce immunoglobulin. ii. T cells iii. NK cells

Arise from progenital cells responsible for the production of granulocytes, monocytes, red blood cells & platelet The differentiation involve : acquisition loss of series of surface antigen defined by monoclonal antibodies.

Antigen Independent Phase

Initial lymphopoiesis takes place in bone marrow Maturation takes places in lymph nodes or thymus
Second cycle of differentiation and proliferation in response to antigen exposure Antibody and Hematopoietic Growth Hormone Production Follows antigen exposure May live for years

Antigen Dependent Phase

Memory Phase

Thrombopoiesis

Production of platelet. Platelet are shed from the cytoplasm of megakaryocytes. Megakaryocytes developed in the bone marrow from the megakaryocyte colony forming units (CFU-Mega) also derived from pluripotential stem cells.

Megakaryocytes maturation : 1. Endomitotic synchronous nuclear replication (nucleus divides, not cell - max 64 segments) 2. Enlarging cytoplasmic volume 3. Cytoplasm become granular 4. Demarcation of membrane of individual platelets Individual platelet or larger fragments are broken off & carried by blood to lungs where the breakdown is completed in pulmonary circulation. They circulate for 6-8 days, then destroyed in spleen or pulmonary vascular bed.

Summary

Haematopoiesis is a process of blood cells production It begins very early in prenatal period and continues till adult The process requires Stem cells Microenvironment Stromal cells Stromal Matrix Other substances Stem cells undergo differentiation & self renewal Progenitor cells undergo differentiation and maturation Mature cells release into blood circulation Most of cells will reenter the tissue phase and undergo apoptosis