Blood Sciences
Blood Sciences
Blood Sciences
Clinical Haematology
Learning Outcomes
• Overview of clinical haematology investigations and
Haematopoiesis
• Describe the stages of red cell production
(Erythropoiesis)
• Describe the regulation of erythropoiesis
• Basic interpretation of red cell parameters
• Terms used to describe red cell morphology
• Abnormalities of the erthyron: Anaemia
Clinical Haematology
• Study of the numbers and morphology of the cellular elements of
the blood
➢ red blood cells (erythrocytes)
➢ white blood cells (leucocytes)
➢ platelets (thrombocytes)
➢ Fibrinogen
➢ Plasma or serum
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Haemopoiesis Model
▪ Most models of haemopoiesis include a number of
compartments
1. Pluripotent stem cell with the capacity for self renewal.
2. Progenitor cells which give rise to different lineages.
3. Progenitor cells gives rise to one lineage.
▪ Note: The above cells are identified by culture techniques
1. Precursor cells
2. Mature end cell compartment
▪ Note the precursor cells and the mature blood cells can
be microscopically identified in bone marrow and blood
samples. The precursor cells will eventually stop dividing
and the maturation process will result in mature cells.
Haemopoiesis
▪ In adult life Haemopoiesis predominantly occurs in the
bone marrow, the soft tissue within the centre of bones
where blood cells develop.
▪ In certain Pathological conditions Haemopoiesis may also
develop in other tissue, including the liver, lymph nodes
and spleen.
▪ Whereas in the foetus as well as the bone marrow
Haemopoiesis occurs in the liver, spleen and lymph nodes
during foetal development.
▪ In the embryo Haemopoiesis occurs in the yolk sac.
Haemopoiesis
▪ In healthy adults in the region of 5-10 x1011 blood cells are
produced in the bone marrow each day.
▪ Haemopoiesis is highly balanced and regulated and relies
upon the following
▪ Growth factors
▪ Cytokines
▪ Environmental factors
▪ Amount of oxygen in the body
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Stem cells and Progenitor cells
▪ Bone marrow
▪ Microscopically Recognisable blood cells in various stages of
development as well as early precursors cells of lineages
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Bone Marrow Trephine Biopsy
Regulation of Haemopoiesis
▪ Haemopoietic growth factors are glycoprotein's hormones
that regulate the proliferation and differentiation of
haemopoietic progenitor cells and the function of mature red
cells.
▪ They regulate the balance between Haemopoiesis and
apoptosis ( ageing cells dying).
▪ The growth factors also react to external stresses such as
infection or blood loss.
▪ The growth factors are produced in different organs in the
body. They bind to surface receptors on target cells ( colony
forming units ) and can trigger replication, differentiation and
functional activation of their target.
Haemopoietic Growth Factors
▪ Haemopoietic Growth Factors or Colony Stimulating Factors (CSF)
are molecules involved in the stimulation, proliferation and
development of progenitor cells.
▪ EPO
▪ G (Granulocyte) CSF
▪ M (Macrophage) CSF
▪ GM-CSF
▪ IL (Interleukin) 3
▪ Thrombopoietin
▪ Stem Cell Factor
▪ VE (vascular endothelial growth factor)
▪ In contrast to classical hormones, haemopoeitic growth factors
and other regulatory cytokines are produced by several cell types
Haemopoietic Growth Factors
Factor Sites of Target Cells produced
Production
EPO Kidney CFU-E Red cells
GM-CSF Fibroblast CFU-GM Granulocyte
Macrophages Macrophages
Bone Marrow Monocytes
Sromal cells
Endothelial Cells
G-CSF As for GM-CSF CFU-GM Granulocytes
(neutrophils)
Thrombopoietin Liver cells CFU-Meg Megakaryocytes
Platelets
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Erythron
• Erythron: all red cells plus all pre-cursor red cell producing
tissues in an animal, including red cells in blood in blood
vessels and sinuses of spleen, liver and bone marrow
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Erythropoietin EPO
▪ Listed are some characteristics of the erythropoietin molecule.
▪ A Sialic acid Glycoprotein with alpha 1 globulin.
▪ 90% is produced in the kidney the remaining 10% is primarily been
produced in the liver.
▪ It is present in small amounts in normal plasma.
▪ The molecular weigh of human EPO is 34 kDa
▪ It is a glycosylated polypeptide of 165 amino acids with a distinct
Electrophoretic pattern
▪ Heat stable
▪ Produced in response to low oxygen tension (PO2). Reduced PO2 leads to
the release of erythropoietin from the kidney.
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Stimulus for Red Cell Production
▪ EPO stimulates RBC production which leads to an
increase in the number of red blood cells and the level of
haemoglobin available to carry oxygen to the tissues.
▪ EPO has been used by trainers to enhance performance
of horses.
▪ Animals living in high altitudes have a higher
concentration of EPO as they require more RBC to pick
up oxygen for the tissues.
▪ Note; a reduction in oxygen in the tissues leads to an
increase in erythropoietin. The erythropoietin acts on the
progenitor cells and the red cell precursors in the bone
marrow to increase the production of the red cells.
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Control of Erythropoiesis
▪ Erythropoietin will influence the process of cell
division in the red cell progenitor cells and the
red cell precursors cells ( the pronormoblasts and
the normoblast).
▪ The erythropoietin will effect receptors that will
signal and influence the level of DNA and mRNA
synthesis within the cell.
▪ Erythropoietin also effects the rate of transfer of
iron to the normoblast and the rate of haem
synthesis.
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Erythropoiesis continued
▪ Normal Erythropoiesis is termed normoblast.
▪ A Myelogram involves the examination the bone marrow
▪ In relation to red cell precursors the normal bone marrow
has the following proportion of immature red cells;
▪ Pro normoblast : 1%
▪ Early normoblast: 1-4%
▪ Inter. normoblast: 20-30 %
▪ Late normoblast: 70%
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Erythropoiesis continued
▪ Myeloid/erythroid ratio: ( MER) its ratio of immature
white cells to immature red cells in the bone marrow.
▪ Normal ratio is 4:1 (white : red)
▪ You have more white cell precursor cells in the marrow
due to the short life span of white cells in the blood.
RBC life span 120 days as opposed to a few days for
most of the white cells with the exception of the
lymphocytes.
▪ In leukaemia and in severe infection the ratio can
increase significantly (20:1 ratio).
▪ In severe blood loss or destruction of red cells you may
get a reduction in the ratio as there may be an increase
in the red cell precursors to boost production of red
cells.
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Erythropoiesis
▪ In a Vitamin B12 and or Folate deficiency, instead of
normoblast, the bone marrow produces Megaloblast.
(Large abnormal red cell precursors ). The bone marrow
is referred to as Megaloblastic.
▪ Hyper refers to over activity or over production of cells
in the bone marrow; Hyperplasic marrow.
▪ Hypo refers to a decrease in activity or underproduction
of the marrow; Hypoplastic marrow
▪ Aplasia or aplastic bone marrows means no production
of cells in the bone marrow.
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Variations from the Normal Maturation
Process in Erythropoiesis
▪ During blood loss or blood destruction, to increase the
production of red cells you get a shortening of the
intermitotic time ( speed of cell division) during the
proliferation and the maturation process.
▪ In iron deficiency anaemia where you get extra cell
division resulting in smaller microcytic cells.
▪ If you omit one or more of the mitotic steps you will a
larger than normal red cell. This occurs when you get in
Megaloblastic anaemia (due to B12 or Folate deficiency)
where you get a Macrocytic cells.
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Measurement of Erythropoieitic Activity
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Measurement of Erythropoieitic Activity
▪ Reticulocyte Count: A high reticulocyte count in the
blood stream would indicate an increase in the rate
of production of red cells.
▪ Blood volume. Red cell mass + Plasma Mass.
▪ Red cell lifespan studies: Use an isotope such as Cr51.
Take a blood sample off a patient and tag it with Cr51.
Inject the cells back into the patient. Measure the
lifespan of the injected cells. Should be 120 days.
▪ Haematinics studies which includes measuring the
levels of B12, Folate, and Iron.
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Bone Marrow Examination
• A bone marrow aspiration and a bone marrow trephine
biopsy are different procedures that often are done
together. The two procedures together are also referred
to as a bone marrow examination.
• Bone marrow is the spongy tissue inside your bigger
bones that produces red blood cells, white blood cells,
and platelets.
• Using a needle, the aspiration draws out a sample of the
liquid portion of your bone marrow. The trephine biopsy
also done with a needle removes a small, more solid part
of the bone marrow.
• The tissue is often taken from the back of your hip bone.
Bone Marrow Biopsy Needles