Hematopoiesis

&

Erythropoiesis
 Objectives – HEMATOPOIESIS

1. Describe the composition of blood

2. Define Hematopoiesis
3. Describe the evolution and formation of blood cells from
embryo to fetus to adult stage
4. Define cytokines, discuss their characteristics and roles in
the differentiation and maturation of hematopoietic
progenitor cells
5. Name anatomic sites involved in hematopoiesis, and the
physiologic function of each
6. Define extramedullary hematopoiesis and the anatomic
sites where this may occur.
 Composition of Blood

Plasma = liquid portion (55% of volume)

Cellular elements = RBCs (45%)
WBCs & platelets (~1%)

Normal adult has 6 L

Comprises 7-8% of total body weight
Hematopoiesis:
the continuous, regulated process of blood cell production that
includes renewal, proliferation, differentiation and maturation.
These processes result in the formation, development and
specialization of all the blood cells released into the circulation
by the bone marrow.
During fetal development, blood cell production occurs in different
parts of the fetus in three phases:
1. The mesoblastic (yolk sack) phase
2. The hepatic (liver) phase
3. The medullary (bone marrow) phase.
 Hematopoiesis General Outline

Stage Begins Where

Mesoblastic 19 days Yolk sac -> AGM (aorta-gonad-mesonephros)
Intravascular sites (in vessels)
Produces embryonic Hb (Gowers, Portland)

Hepatic 5-7 wks Major site is liver

Also spleen, kidney, thymus, lymph nodes
Clusters of recognizable cells, blasts
Hb – predominantly Hb F, start of Hb A

Medullary Late Medullary = Inner part of bone

(adult) 4th mo At 6 mos, bone marrow takes over as primary site
All cell types produced – in BM, also lymph nodes,
spleen, liver, thymus
Lymphoid tissues – primarily BM and thymus, also
spleen, lymph nodes and other lymph tissues
Hematopoiesis General Outline
 Summary

1. Mesoblastic or Yolk sac phase: 2 – 10 weeks

2. Hepatic or Liver phase: 1 – 9 months, peaks at about 5 months
3. Medullary or Blood Marrow phase: 4 months on. By end of sixth
month, bone marrow is the primary site of hematopoiesis.
4. Adult phase: All hematopoietic elements have developed.

months / years
 Fetal Hematopoiesis
1. Yolk sack phase:

♥ In humans begins at about the 19th day of embryonic development.

♥ Primitive erythroblasts originate from the lining of the embryonic

yolk sack.

♥ Erythroblasts produce some primitive forms of hemoglobin but do

not contribute significantly to definitive hematopoiesis.
2. Liver phase:
♥ At 4-5 weeks liver becomes erythropoietic center and remains so
until one to two weeks after birth.
♥ During fetal development the developing spleen, kidney, thymus
and lymph nodes also contribute to blood cell production.
♥ Production of megakayocytes begins in the hepatic phase.
3. Medullary phase:

♥ By five months, hematopoiesis begins in the bone marrow.

♥ By six months the bone marrow is the primary site of blood cell
production.

♥ Various stages of maturation in all cell lines can be seen.

 Adult Hematopoiesis
♥ The adult the bone marrow contains developing erythroid,
myeloid, megakaryocitic and lymphoid cells.

♥ Lymphoid cell development is in the marrow, thymus, spleen and

Lymph nodes.
 Body places of Hematopoiesis
1. Bone Marrow:
• One of the largest organs in the body.
• Honeycomb areas within large bones contain red and yellow marrow.
• Red marrow: a complex arrangement of hemopoietic elements.
• Yellow marrow: mainly fatty tissue.
In adults, active marrow is mainly
in the large, flat bones and the ends
of some long bones.
Bone Marrow cont…

A. Major site of blood cell production: 6 months to adult

B. One of the largest organs of the body
C. Located in the cavities of the cortical bones
D. Network of 3-D honeycomb matrix (structural support)
E. Advanced system of vessels regulates flow of particles

Marrow
A. Red – active; developing blood cells
B. Yellow – inactive; fat cells, undifferentiated mesenchymal &
macrophages
C. Contains cells; endothelial, macrophages, lymphs, osteoblasts,
osteoclasts, fibroblasts
D. Contains substances; fibronectin, collagen, proteoglycans,
etc.
Bone marrow cont..
Hematopoiesis cont…

2. Liver:

A. The center of hematopoiesis the hepatic phase during fetal

development.

B. The liver can become hemopoietically active if needed in adults

as in the case of bone marrow disease or damage.

C. The liver is a source of metabolic support for the blood

production and processes such as bilirubin metabolism.
Liver cont…

http://www.medicinenet.com/liver_disease/article.htm
Hematopoiesis cont…

3. Spleen:
A. The largest lymphoid organ.
B. Normally contains about 350 ml of blood.
C. The spleen is divided into discrete regions containing red
pulp, white pulp and a marginal zone.
White pulp contains germinal centers with lymphocytes,
macrophages and dendritic cells (immune cells not in brain).
Red pulp has a structure that allows it to serve as a
sponge like filter.
D. Indiscriminate filter of circulation
Culling – damaged, degraded cells are phagocytized
Pitting – macrophages remove inclusions or damaged
surface membrane
Spleen cont….
E. Recycles red cells
F. Storage site for platelets (30% of total)
G. Immune defense against bacterial infections
H. The spleen stores platelets – approximately 30% of a normal
individual’s platelets are held in the spleen. In addition,
30% of WBC also stored in the spleen.
I. It can be a source of extramedullary hematopoiesis when
needed.
J. Splenomegaly can result from many causes such as, chronic
leukemias, genetic RBC defects, malaria, Hodgkin disease,
myeloproloferative disorders and thalassemias.
Spleen cont….

Slideplayer.com
https://vimeo.com/6805886
Hematopoiesis cont…

4. Lymph Nodes
• Occur in groups or chains along the lymphatic vessels.
• Have an outer cortex and inner medulla; both have complex
structure and contain lymphocytes and macrophages.
• Produce lymphocytes from germinal centers, process
immunoglobulins and filter particulate matter and bacteria
entering from the lymph.
Lymph Nodes cont…
•
 Lymphatic organs, parallel circulatory system
 Lymph – fluid part of blood, traverses connective tissues,
has low protein concentration, no RBCs

 Functions:
 Immune defense against pathogens
 Lymphocyte proliferation
 Initiate specific immune response to foreign antigens
(germinal center).
 Filter particulate matter, debris, bacteria in lymph
Hematopoiesis cont…

5. Thymus
 Well developed at birth, atrophies by old age.
 12 – 15 g at birth, 30 – 40 g at puberty, gradually smaller as age
increases.
 Located in the upper mediastinum and has cortex and medulla
like other lymphoid tissue.
 Site of production of T-lymphocytes from progenitor lymphoid
cells that have migrated fro the bone marrow.
Hematopoiesis cont…

Thymus

McKenzie, S., Williams, J. Clinical Laboratory Hematology, 3rd ed, 2015

Hematopoiesis cont…

Thymus

 In utero, until birth – stores primitive lymph cells and

macrophages, also mesenchymal, reticular, epithelial,
dendritic

 Adults – progenitor cells from BM travel to the thymus to

‘mature’(obtain CD4 and CD8 surface antigens)
Blood Cell Origination
Blood Cell Origination
HSC – Hematopoietic Stem Cell, 3 fates:
1. Self-Renewal
2. Differentiation
3. Apoptosis
Committed Progenitor Cells
CLP – common lymphoid progenitor
CMP – common myeloid progenitor (CFU-GEMM)
Hematopoiesis cont…

1. Cytokines and growth factors affect differentiation of

stem cells. (Kine - Greek: movement).

2. Cytokines include interleukins, colony stimulating factors,

& interferons.

3. Some cytokines influence stem cells with multilinear

potential, some are lineage specific, some work in
combination with other cytokines.

4. Cytokines are necessary to prevent premature cell death

(apoptosis).

5. Cytokines provide new tools in the treatment for diseases

such as bone marrow malignancies, leukemia and aplastic
anemias.
Interleukins:
Definition = glycoproteins that regulate proliferation,
differentiation, and maturation of hematopoietic precursors

Characteristics
1. Proteins that exhibit multiple biological activities (regulation
of autoimmune and inflammatory reactions, and
hematopoiesis)
2. Interact synergistically with other cytokines
3. Have amplification potential
4. Effective at low concentrations
Main specific proteins
EPO (Erythro-) prod by kidney, much lesser amount in liver
TPO (Thrombo-) prod mainly in liver
Hematopoiesis cont…
Specific cell development:

1. Erythropoiesis
2. Leukopoiesis
3. Megakaryopoiesis
Hematopoiesis cont….

• When blood cell production occurs outside the marrow it is

termed extramedullary.
• The marrow microenvironment is essential for stem cell differentiation
and proliferation.
• Monophyletic theory holds that all types of blood cells originate from a
common stem cell.
• Maturation – as red blood cells mature, they generally become smaller,
as does the nucleus.
• Nucleoli are lost and nuclear chromatin becomes denser.
• Cytoplasm becomes less basophilic.
 Erythrocyte (red blood cell) Production and
Destruction

1. Erythrocytes (RBCs) develop from progenitors

dedicated to the erythroid line.

2. These are the BFU-E (burst forming unit-erythroid)

and CFU-E (colony forming unit-erythroid) types.

3. They give rise to the morphologically identifiable

cells of the erythroid line.
 RBC Development
A. Definition of Erythropoiesis = RBCs develop from
committed erythroid progenitor cells

B. Cells mature in organized, morphologically

identifiable cells

C. Nucleus – becomes condensed, ultimately extruded

D. Cytoplasm – staining color changes from blue

(ribosomes) to salmon-pink (hemoglobin)

E. 18-21 days for full maturation

F. Life span of mature RBC is 120 days

 RBC Development
A. Cell diameter decreases, cytoplasm changes from blue
to pink

B. Nuclear diameter decreased, color changes from

purplish red to very dark purple

C. Nuclear chromatin becomes coarser, clumped, condensed

D. Composite illustration of all changes

 ERYTHROPOIESIS

Ref: Keohane, 5th ed

.
Erythrocyte (red blood cell) Production and Destruction cont…

In order from least to most mature:

1. Pronormoblast (Rubriblast)

2. Basophilic normoblast (Prorubricyte)

3. Polychromatophilic normoblast (Rubricyte)

4. Orthochromic normoblast (Metarubricyte)

5. Reticulocyte (Polychromatophilic erythrocyte)

6. Erythrocyte
Erythropoiesis
1. Pronormoblast
The nucleus takes up most of the cell
One or two nucleoli
Fine chromatin pattern.
Cytoplasm blue
Golgi complex may be visible
 2. Basophilic normoblast
Some chromatin clumping
Nucleus more compact
Cytoplasm deep purple-red (basophilic)

,,
3. Polychromatic normoblast
Nucleus smaller
chromatin more condensed
no nucleoli
Hemoglobin is apparent in the blue cytoplasm
(polychromatophilic).
4. Orthochromic normoblast
Nucleus smaller and very condensed
Cytoplasm has hemoglobin color throughout with a
bluish tint.
5. Reticulocyte
No nucleus, but nuclear remnants stain with supravital
stains.
Cytoplasm has bluish tint and the cell is slightly larger
than mature RBCs.
6. Mature erythrocyte
Erythrocyte Production and Destruction cont….

As erythroid cells mature, the nuclei condense and are ejected

from the cell, producing the reticulocyte.
Wright stained cytoplasm changes from blue to pink as numerous

organelles of less mature cells are replaced by hemoglobin.

Changes in the nuclei and stained cytoplasm identify each stage.

RBCs mature in the blood marrow in approximately two

weeks.

They are identifiable as RBC precursors for only the last third of

that time.

RBCs have a half life or last about 120 days.

Erythrocyte Production and Destruction cont…

• Hypoxia of the peripheral blood causes peritubular cells of the

kidney to secrete erythropoetin (EPO), the hormone that
stimulates the production of RBCs.

Erythropoetin EPO:
• Prevents cell death of RBC precursors
• Reduces the number of times precursors divide before
beginning maturation
• Speeds up cell division
Erythrocyte Production and Destruction cont…

• Apoptosis is the mechanism by which excessive production of RBCs

is controlled.
• Young precursors have a death receptor, Fas, and older precursors
have an FasL ligand.
• As older RBC precursors mature more slowly, they suppress the
production of younger cells.
Erythrocyte Production and Destruction cont….

• EPO rescues cells by stimulating the production of molecules that

counteract the actions of Fas and FasL

• Survival of RBC precursors in the bone marrow depends on

cytokines and fibronectin which are produced by macrophages.

• As RBCs mature, they lose fibronectin receptors and can leave the
bone marrow.

• Mature RBCs leave the marrow through pores in the endothelial

cells in the venous sinus.
Erythrocyte Production and Destruction cont…

• Mature RBCs that can not regenerate cellular constituents and

eventually loose the ability to maintain their shape, are trapped in
the spleen and engulfed by macrophages.
This is the main mechanism for the destruction of old RBCs.

• Destruction in the circulating blood, or intravascular destruction,

occurs when circulating RBCs are damaged in various ways, and is
a minor route for RBC destruction.
 Questions
1. Blood cells, red and white, originate in the
bone marrow. Which cell type matures
outside of the bone marrow?

2. Which organ is the storage site for platelets?