Haematopoiesis:

formation of blood cells

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 Haematopoiesis
• aka haemopoiesis
• and nb if US spelling, hemo-

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 Haematopoiesis
• This is the process by which ALL blood
cells (= haematological system) are
produced
– platelets, red blood cells, leukocytes of all
sorts
– (AND probably also endothelium).

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 Haematopoiesis
• Haematopoiesis is a highly organised
differentiation process involving the
ordered expression of different sets of
genes
• It is controlled by factors in the
environment of the developing blood cell
– the bone marrow in adults
• just like any other developmental
programme.
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 The haematopoietic stem cell

• Mature blood cells in healthy individuals

mostly have short lifetimes (exception:
lymphocytes) and are constantly
regenerated in the bone marrow.
– we make 5 x 1011 blood cells daily
• This is accelerated when there is
haematological stress
– e.g. infection, need more leukocytes
– e.g. high altitude, need more red cells.

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 Haematopoiesis
• Haematopoiesis begins at a very early
stage in embryonic development, at about
3 weeks in the human.
• At that time the cells in the embryo
separate into 2 sets, one generating the
embryo proper & all the tissues of the
adult, the other forming the YOLK SAC
(aka vitelline sac) which is the site where
blood cells and blood vessels are first
formed. 6
 Haematopoiesis
• The yolk sac is an ovoid structure joined to
the embryo by a stalk.
• It contains mesoderm derived cells
– “haemangioblasts”
• which differentiate to form (nucleated) red
blood cells and endothelial cells which
generate a capillary system (“plexus”)
within the yolk sac.
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 Haematopoiesis
• At the same time the heart and aorta start
to form: these join up with the capillary
plexus and the erythrocytes start to
circulate.

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Human foetus
about 5 weeks gestation
YS = yolk sac Ao = aorta
Square = location of limb bud

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Definitive haematopoiesis

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 Haematopoiesis
• At a later stage of embryogenesis (after
about week 6 in humans) haematopoiesis
occurs mainly in the liver and at birth shifts
to the bone marrow (BM).
• This haematopoiesis is described as
definitive unlike the early primitive
haematopoiesis occurring in the yolk sac.
• Now, the entire range of blood cells found
in the adult are produced.
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 Haematopoiesis
• The BM is in effect a highly specialised
tissue comprising a range of cells
– some of which (haematopoietic cells) form
blood cells
– others (stromal cells) provide a support
functions for the haematopoietic cells,
providing the specialised environment needed
for haematopoiesis to occur.
– Yet other cells (osteoblasts and osteoclasts)
are concerned with producing the bone itself.
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 Haematopoiesis
(Bear in mind that cells of the immune
system undergo further proliferation and
differentiation in the periphery – especially
in secondary lymphoid tissue – during
immune responses: the purpose of
haematopoiesis is to generate cells which
are capable of responding to pathogens.)

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 BONES:
where haematopoiesis occurs

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dem bones –
more than a framework
to hang muscles from

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 bones
• Source of skeletal rigidity
• reservoir of Ca2+ and PO43-
• haematopoietic organs.

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 bones
• Bone is a specialised form of connective
tissue (mesodermal/mesenchymal origin)
with an extracellular matrix (ECM) which is
rigid
http://www.emedicine.com/orthoped/TOPIC403.HTM

• Rigid outer layer of dense compact (aka

cortical) bone, 70% hydroxyapatite
(hydrated calcium phosphate)
• Inner core of much less dense spongy
bone (aka cancellous, trabecular).
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Long bone

(location of BM)

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Schematic of cross section of long bone

contains blood
vessels nerves
and lymphatics

joins together
http://en.wikipedia.org/wiki/Bone Haversian canals

The osteon is the unit structure of compact bone made up of

concentric mineralised lamellae around a central (Haversian) canal.
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Histological section of compact bone showing Haversian and
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Volkman’s canals.
 bones
• The bone marrow (BM) is within the
medullary cavity especially of long bones.
• As mentioned it is intensely cellular
because that is where blood cells are
produced.

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Histological section of bone marrow

blood vessels

bone trabeculae

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trabecula = strand (here, of bone)
 bones
• Bones always start (in development) from
cartilage: this is converted to bone by
deposition of hydroxyapatite (a form of
calcium phosphate) by osteoblasts.
• But throughout life, bone tissue is renewed
– this is due to a balance between degradation
due to osteoclasts and production due to
osteoblasts.

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 bones
• Osteoblasts (a type of BM stromal cell) are
responsible for laying down a collagen
matrix (osteoid) & they subsequently
mineralise this with hydroxyapatite.
• They are activated by growth factors e.g.
bone morphogenic protein (BMP) & certain
steroid hormones e.g. oestrogens.

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 bones
• Osteoclasts (derived from haematopoietic
cells) resorb bone.
• They are activated by certain cytokines
e.g. IL-6.
• They secrete acids which dissolve the
hydroxyapatite and proteases which
degrade the collagen.

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 bones
• This balance between production and
degradation can be disturbed in some
diseases where the osteoclasts get the
upper hand
– e.g. osteoporosis due to low levels of
oestrogen, myeloma due to high levels of IL-6.

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Osteolytic lesions in
a case of myeloma.

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The haematopoietic stem cell:
source of all blood cells

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 The haematopoietic stem cell

• Haematopoietic stem cells (HSCs) are the

parental cell type for ALL blood cells
– and probably also endothelium.
• HSCs are (usually) present only in the
bone marrow
– also present in cord blood and in (adult)
peripheral blood after treatment with
“mobilising” cytokines.

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 The haematopoietic stem cell

• The HSC is (probably) able to divide

indefinitely, or at least a very large number
of times
– experiments in 1960s in which mice were
irradiated or treated with drugs to destroy their
bone marrow showed that transplantation of
very small numbers of bone marrow (BM)
cells could “reconstitute” the mouse
– serial transplant could be done.
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 The haematopoietic stem cell

• Experiments in the 1970s showed that all

blood cell types could be derived from
single BM cells.

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 The haematopoietic stem cell

• These depended on irradiating the BM

cells used to reconstitute the mice with
SMALL doses of radiation, not enough to
kill the cells but enough to cause minor
chromosome alterations (“chromosomal
markers”) in rare cells
– each marker unique;
– all descendents of a marked cell (a CLONE)
have the same maker.
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 The haematopoietic stem cell

• In some individual transplanted mice there

could be found cells of all leukocyte types
bearing the same marker
• hence a single pluripotent stem cell present
in the transplant was capable of giving rise to
all lineages.
– (Probably not totipotent i.e. cannot make all body
cells.)

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 The haematopoietic stem cell

• In other mice, ALL myeloid but NOT

lymphoid cells had the same marker
• And in yet others ALL lymphoid but NOT
myeloid cells had the same marker.

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 The haematopoietic stem cell

• This implied that there were also stem

cells of more restricted potency
– capable of producing EITHER myeloid OR
lymphoid cells
– these are now known as common lymphoid or
myeloid progenitor cells
– CLP & CMP.

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 The haematopoietic stem cell
• Hence:
CLP
(common lymphoid Lymphocytes
progenitor)
HSC
CMP Granulocytes
(common myeloid erythrocytes
progenitor) thrombocytes

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 The haematopoietic stem cell

• Stem cell self renewal:

– evidently since mature leukocytes die the
HSC must be able to proliferate at least over
the lifetime of the individual.
– This proliferation must be assymetric to
generate a progenitor cell (CMP or CLP)
which has more limited differentiation capacity
AND replace the stem cell.

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 The haematopoietic stem cell

• CMPs and CLPs are transient amplifying

(TA) cells & proliferate PRIOR TO further
differentiation (i.e. before they become
granulocytes or whatever)
– this is a limited proliferation, increasing cell
numbers.

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 The haematopoietic stem cell

proliferation of CMP CMP

progenitors to CMP
amplify numbers CMP CMP
CMP
CMP CMP
(or CLP) CMP
CMP
HSC CMP

CMP
asymmetric division to generate
progenitor cell & replace HSC 39
 The haematopoietic stem cell

• What is the HSC?

– A rare (<0.1%) cell present in bone marrow
expressing the antigen CD34.
– We now know that CD34+ cells are the
essential cell type for BONE MARROW
TRANSPLANTATION.
– Other cells express CD34 (e.g. endothelial
cells) so it us not a unique marker for HSCs.

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 The haematopoietic stem cell

• It is argued that stem cells require a

specialised environment to maintain the
“stem-ness” of the stem cell:
– the stem cell niche.
• HSCs are found in two locations in BM:
– the endosteum (boundary between solid bone
& marrow) associated with osteoblasts;
– the perivascular region around vascular
sinusoids (large blood vessels with thin-walled
comprised of fenestrated endothelium). 41
 The haematopoietic stem cell

• Hence “endosteal” and “vascular” niches.

• Do they have different roles?
– It has been suggested that the endosteal
niche contains long-term, slowly dividing
HSCs which maintain the vascular niche
HSCs.
– In the vascular niche the HSCs are more
actively dividing and progenitor cells are
produced.

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HSC niches in the BM
endosteal niche vascular niche

osteoblast endothelium

sinus

CMP

CMP

HSC
HSC

CLP 43
endosteum
 The haematopoietic stem cell

• It is not clear what molecules are involved

in defining the stem cell niches
– presumably adhesion molecules & diffusible
factors produced by the non-HSC (“stromal”)
cells – osteoblasts, endothelial cells etc –
within the niche.

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Li, Z. & Li, L. (2008) Understanding hematopoietic stem-cell
microenvironments. Trends in Biochemical Sciences 31:589-505
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Say no more
Progenitor cell growth and
differentiation

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 growth and differentiation

• As the HSC divides, one cell leaves the

niche and becomes a progenitor cell, the
other stays put
– asymmetric division in space as well as in
kind

niche containing niche containing niche containing stem cell

stem cell dividing stem cell and progenitor cell
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outside niche
 growth and differentiation

• Evidently the progenitor cell is now in a

different environment.
– Growth factors drive its growth and then
differentiation.
– (Perhaps the job of the stem cell niche is to
slow cell division and block differentiation.)
– The different phases of differentiation and the
growth factors required have been largely
worked out by cell culture experiments.
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 growth and differentiation

• In vitro culture of bone marrow cells

– culture of BM cells is often done suspended in
semi-solid media so that the progeny of a
single cell can be seen as a colony.

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Haematopoietic colonies in semi-solid medium

other blood cells

erythrocytes

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 growth and differentiation

• Alternatively, cells can be grown in liquid

culture
– this requires the presence both of
haematopoietic cells and BM stromal cells to
recreate something like the stem cell niche
– even so cell growth is very inefficient.

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 growth and differentiation
• In both cases added growth factors &
cytokines are needed to drive both
proliferation and differentiation of cells.

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 growth and differentiation
• There are many of these, some more-or-
less specific for different cell types
– e.g. Granulocyte/monocyte colony colony
stimulating factor (GM-CSF), erythropoeitin
(EPO)
– NB the term CSF.
• Others with much broader activities
– e.g. IL-3 which will stimulate growth of most
haematopoietic cell types.
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 growth and differentiation
• Growth factors/cytokines act both in a
paracrine fashion
– they diffuse from the producer cell to the
responder cell
• and in a juxtacrine fashion
– they remain on the cell surface of the
producer cell and so the responder cell must
be juxtaposed – touching.

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 growth and differentiation
• And they are produced both within the BM
and from sources outside the BM
– e.g. GM-CSF from lymphocytes, EPO from
kidney.
• Infection can massively increase the
amount of growth factors present because
of activation of a range of leukocytes at
the site of infection
– hence increased haematopoiesis.
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 growth and differentiation
• Adhesion interactions especially via
integrins are also important in
haematopoiesis
• not just to drive growth & differentiation but
also to prevent apoptosis.

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erythrocytes & platelets

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 erythrocytes & platelets
• The early stages for production of both
these cells pass through a common
pathway generating a
“megakaryocyte/erthrocyte precursor”
(MEP) from the CMP: they then split.

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 erythrocytes & platelets
• erythropoietin (EPO: aka epoietin)
produced by kidneys specifically
stimulates erythropoiesis
– EPO production stimulated by hypoxia
– EPO binds to erythrocytes so haemorrhage or
anaemia results in increased levels of EPO
– these lead to stimulation of erythropoiesis.
– (Other non-specific GFs/cytokines needed.)

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Simplified erythropoiesis
bone marrow circulation
pro-
MEP erythroblast erythroblast* reticulocyte** erythrocyte

EP EP
EP EP

dividing cells committed to non-dividing cells

erythroid lineage: requires EPO (etc)

Hb synthesis
*The suffix “-blast” indicates a large, proliferating cell. There
are several different kinds of erythroblast.
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**The reticulocyte nucleus is condensed and inactive.
 erythrocytes & platelets
• Erythrocyte clearance:
– removed by liver and spleen after 120 days
– as the erythrocyte ages surface proteins
particularly “band 3” are progressively
oxidised
– this provides a target for phagocytosis by
macrophages lining liver & spleen sinuses.

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 erythrocytes & platelets
• Generation of platelets (thrombopoiesis):
– stimulated by thrombopoietin (TPO) & other
non-specific growth factors.
– TPO produced constitutively mostly by liver.
– Inflammation can double production by liver
via cytokine IL-6.
– In thrombocytopenia (reduced platelets) BM
stromal cells also produce TPO.
– Platelets have TPO receptors and so remove
TPO from circulation (-ve feedback).
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Thrombopoiesis
bone marrow circulation
megakaryocyte platelets
MEP

requires TPO (etc)

Platelets (1,000s) bud off surface of megakaryocyte.

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 erythrocytes & platelets
• Platelet clearance:
– removed by liver and spleen after ~ 7 days .

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Granulocytes and monocytes

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 granulocytes and monocytes
• Granulocytes (basophils, mast cells,
eosinophils & neutrophils) and monocytes
share early steps in differentiation via
“granulocyte/monocyte precursor” (GMP)
derived from the CMP.
• Formation of GMP from CMP is driven by
granulocyte/monocyte colony stimulating
factor (GM-CSF) (etc).

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 granulocytes and monocytes
• Formation of specific cell types is driven
by other growth factors, all of which are
produced constitutively and are induced
e.g. by inflammation.

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Granulopoiesis neutrophil

neutrophil
bone marrow tissues
circulation

GMP G-CSF
mast cell
SCF*
basophil
IL-5

M-CSF eosinophil

eosinophil
*SCF = Stem cell factor.
Note that some cell types macrophage
differentiate further in the
tissues. 68
monocyte
 granulocytes and monocytes
• Most of these cells are relatively short
lived and are lost by apoptosis
– the fragments of the dead cells are
phagocytosed by cells lining sinuses of spleen
& BM
– or, in the tissues, by epithelial cells.

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lymphocytes

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 lymphocytes
• Two pathways:
– T cells
– B cells and NK cells
• Both from CLP
• Initial stages in BM: B & NK cells complete
maturation in BM, immature T cells
migrtae to thymus and mature there.

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T lymphopoiesis
circulation & 2ry
bone marrow thymus lymphoid tissue
CLP DN DP mature T (SP)

IL-7R
CD4/8 +ve
CD25 +ve
α chain re-
arrangement
β chain re-
arrangement 72
B lymphopoiesis
circulation & 2ry
bone marrow lymphoid tissue
CLP pro-B pre-B mature B

IL-7R
CD19 +ve
IgM +ve
H chain re-
arrangement
L chain re-
arrangement 73
lymphopoiesis abbreviations

DN = negative for both CD4, 8; DP = positive for both; SP =

positive for one or other
IL-7R = IL-7 receptor
CD4/8 +ve = expressing one, other or both of these T cell markers
CD25 +ve = expressing IL-2 receptor
α chain, β chain re-arrangement = formation of T cell receptor
IgM =ve = synthesising IgM
CD19 +ve = expressing standard B cell marker
H chain, L chain re-arrangement = formation of mature Ig genes

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 lymphocytes
• The process of lymphopoiesis is driven
initially by the growth factor IL-7
– hence the importance of its receptor.
• How the different lymphoid lineages are
established & what factors influence this is
complex and ill-understood
– by me at any rate.

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 lymphocytes
• Lymphocytes are long lived cells unless
they are activated (by cognate antigen).
• They then become immune effector cells
– most of these will die after a few days by
apoptosis
– some will become memory cells which are
very long lived (decades).

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 Conclusions
• Haematopoiesis is a complex business but
the outlines are pretty clear.

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