HEMATOPOYESIS

MEMB
 Classic example of the biological principle that cells
have specialized functions and that their structures are

RBC specific for those functions

Function: to carry oxygen from the lung to the tissues,

where the oxygen is released.
erythrocyt
e This is accomplished by the attachment of the O2 to
hemoglobin (HGB), the major cytoplasmic component
of mature RBCs.

Diminished The role of the RBC in returning CO2 to the lungs and
oxygen-carrying buffering the pH of the blood is important but is quite
secondary to its oxygen-carrying function.
capacity of the
blood, called The mechanisms controlling development, production,
and normal destruction of RBCs are fine-tuned to avoid
anemia. interruptions in oxygen delivery, even under adverse
conditions such as blood loss with hemorrhage.
• The mammalian erythrocyte is
unique among animal cells in
having no nucleus in its mature,
functional state.

• While amphibians and birds possess

RBCs, their nonmammalian RBCs
retain the nuclei throughout the
cells’ lives.

• The implications of this unique

mammalian adaptation are
significant for cell function and life
span.
 CELLULAR MODEL OF ERYTHROID DIFFERENTIATION

Although the two classes of committed erythroid progenitors (BFU-E and

CFU-E) appear distinct from each other, in reality progenitor cells
constitute a continuum, with graded changes in their properties.
 CMP in turn give rise to
Multipotent stem cells generate cellular
granulocyte/macrophage progenitor
compartments defined on the basis of their
(GMP) and megakaryocyte erythroid
antigenic profile and restricted toward the
progenitor (MEP), which probably
myeloid differentiation pathway–defined
correspond to the burst-forming unit–
common myeloid progenitor (CMP)
erythroid (BFU-E)

Lastly, MEP generate cells capable of

Each erythroid colony or burst
uniline age differentiation toward either
derives from one BFU-E or
the megakaryocytic (colony-forming unit-
CFU-E, respectively. CFU-GM,
megakaryocyte [CFU-Mk] or the
Colony-forming unit–
erythroid pathway (colony-forming unit–
granulocyte-macrophage
erythroid [CFU-E]).

A multilineage colony-forming unit (CFU-GEMM) or common myeloid progenitor,

and the most primitive BFU-E have physical and functional properties that are
shared by both pluripotent stem cells and progenitor cells committed to
nonerythroid lineages.
 The first, more primitive class consists
of the burst-forming unit–erythroid (BFU-
E), named for the ability to give rise to
multi clustered colonies (erythroid
bursts) of hemoglobin-containing cells.

BFU-E represent the earliest progenitors

committed exclusively to erythroid
differentiation and a quiescent reserve,
with only 10% to 20% in cycle at any
given time.

They have a limited self-renewal

capacity; at least a subset of BFU-E is
capable of generating secondary bursts
upon replating.
 A second, more differentiated class of progenitors
consists of the colony-forming unit–erythroid (CFU-E).

Most (60% to 80%) of these progenitors already are in cycle and

thus proliferate immediately after initiation of culture, forming
erythroid colonies within 7 days

Because CFU-E are more differentiated than BFU-E, they require

fewer divisions to generate colonies of hemoglobinized cells, and the
colonies are small (8 to 64 cells per colony).
BFU-E (burst forming unit-erythroid) stem cell, derivada de la célula pluripotencial,
que da origen a granulocitos, monocitos, eritroblastos, megacariocitos.

A medida que el proceso de maduracion progresa se desarrolla la CFU-E, sumamente

sensible a la acción de la eritropoyetina.
 1) stromal cells (e.g., fibroblasts,
endothelial cells, mesenchymal stem
cells and their diverse descendant
progeny)

The bone marrow

microenvironment
consists of three broad
components:

2) accessory cells (monocytes, 3) extracelular matrix (a

macrophages, megakaryocytes, protein-carbohydrate
T cells) scaffold)
 In the bone marrow, described
two distinct hematopoietic
niches for hematopoietic stem
cells /progenitor cells:

The “endosteal” niche and the

Reciprocal communication of
hematopoietic stem cells with
cells/matrix in their bone
marrow niche ensures both
their quiescent state and self-
renewal dynamics
“endothelial or vascular niche”
within the medulla, recent studies
suggest that this distinction may
be artificial, because both cellular
structures can be intimately
associated in trabecular bone

Thus mesenchymal stem cell–derived

osteoprogenitor cells and stromal reticular cells
(nestin+ or leptinR+) are intimately associated
with sinusoidal endothelium and seem to be
pivotal organizers of the bone marrow niche
• The nucleus-to-cytoplasm (N:C) ratio is a morphologic feature used
to identify and stage red blood cell and white blood cell precursors.

• If the areas of each are approximately equal, the N:C ratio is 1:1.

• If the nucleus takes up less than 50% of the area of the cell, the
proportion of nucleus is lower, and the ratio is lower (e.g., 1:5 or
less than 1).

• If the nucleus takes up more than 50% of the area of the cell, the
ratio is higher (e.g., 3:1 or 3).

• In the red blood cell line, the proportion of nucleus shrinks as the
cell matures and the cytoplasm increases proportionately, although
the overall cell diameter grows smaller. In short, the N:C ratio
decreases.
 Normoblastic Series: Summary of Stage
Morphology

(reticulocyte)
Pro normoblasto
 Erythrokinetics
• Term describing the dynamics of RBC production and
destruction.

• To understand erythrokinetics, it is helpful to appreciate

the concept of the erythron.

• This discussion of erythrokinetics begins by looking at

the erythrocytes in the bone marrow and the factors
that affect their numbers, their progressive
development, and their ultimate release into the blood.
 Eritrón
Por definición, comprende todos los estadios de los eritrocitos, en diferentes áreas del
cuerpo:
- Normoblastos en desarrollo en MO y
- Eritrocitos maduros circulantes en sangre periférica y los espacios vasculares en
órganos específicos como el bazo

El eritrón se distingue de la masa de glóbulos rojos

El eritrón es la totalidad de las células eritroides en el cuerpo, mientras que la masa de

glóbulos rojos se refiere solo a las células en circulación

Cuando se emplea el término eritrón, está implícito el concepto de un tejido funcional

unificado
Mature erythrocytes and one lymphocyte, peripheral blood (Wright
stain, 31000). B, Scanning electron micrograph of mature erythrocytes
Ciclo de retroalimentación entre la médula
eritroide y los tejidos del organismo
Hypoxia—the Stimulus to Red Blood Cell
Production
• The primary oxygen-sensing system of the body is located in
peritubular fibroblasts of the kidney.

• Hypoxia, too little tissue oxygen, is detected by the peritubular

cells, which produce erythropoietin (EPO), the major stimulatory
cytokine for RBCs.

• Under normal circumstances, the amount of EPO produced

fluctuates very little, maintaining a level of RBC production that
• is sufficient to replace the approximately 1% of RBCs that normally
die each day.

• When there is hemorrhage, increased RBC destruction, or other

factors that diminish the oxygen-carrying capacity of the blood, the
production of EPO is increased.
• Hypoxia increases EPO production in peritubular cells mainly by
transcriptional regulation.

• The EPO gene has a hypoxia-sensitive region (enhancer) in its

regulatory component.

• When oxygen tension in the cell decreases, hypoxiainducible

factor-1, a transcription factor, is assembled in the cytoplasm,
migrates to the nucleus, and interacts with the enhancer of the
gene.

• This results in transcription of more EPO messenger RNA

molecules, and production of more EPO.
 box
• The location of the body’s hypoxia sensor in the kidney is practical,1 because the kidney
receives approximately 20% of the cardiac output with little loss of oxygen from the
levels leaving the heart.

• The location provides early detection when oxygen levels decline.

• Making the hypoxia sensor the cell that is able to stimulate red blood cell (RBC)
production also is practical, because regardless of the cause of hypoxia, having more
RBCs should help to overcome it.

• The hypoxia might result from decreased RBC numbers, as with hemorrhage,
decreased RBC number, however, is only one cause of hypoxia.

• Another cause is the failure of each RBC to carry as much oxygen as it should. This can
occur because the hemoglobin is defective or because there is not enough hemoglobin
in each cell.
 box
• The hypoxia may be unrelated to the RBCs in any way; poor lung function resulting in
diminished oxygenation of existing RBCs is an example.

• The kidney’s hypoxia sensor cannot know why there is hypoxia, but it does not
matter.
• Even when there are plenty of RBCs compared with the reference interval, if
there is still hypoxia, stimulation of RBC production is warranted because the
numbers present are not meeting the oxygen need.

• An elevation of RBC numbers above the reference interval, erythrocytosis, is

seen in conditions such as lung disease and cardiac disease in which the blood
is not being well oxygenated.

• Newborns have higher numbers of RBCs because the fetal hemoglobin in their
cells does not unload oxygen to the tissues readily, so newborns are slightly
hypoxic compared with adults. To compensate, they make more RBCs.
 Sitio probable de la
síntesis en células
intersticiales peritubulares
localizada en el riñón

ERITROPOYETINA

Al unirse con los receptores Se estimula por la

de superficie de los hipoxia tisular y es
precursores eritroides, capaz de aumentar la
estimula la síntesis de RNA y masa de glóbulos rojos
de cAMP
Unida a receptors de membrana específicos (EPOR), puede estimular a las
CFU-E y controlar la producción de eritrocitos mediante:

1. La regulación de las tres fases de división-reducción de la producción

normoblástica

2. El control de la tasa de producción por acortamiento del tiempo

del proceso de división y/o de maduración

3. El aumento de la velocidad del ciclo de la pentosa fosfato

4. Su acción en las paredes de los senos de la médula ósea, donde

favorece la salida de eritrocitos maduros mediante brechas pequeñas
del endotelio hacia los sinusoides

6. Aumento de la velocidad de síntesis de la hemoglobina mediante la

transferencia de hierro de la transferrina a los precursores eritroides en
desarrollo
 It is produced A heme-containing protein
EPO, a 35-kd glycoprotein, is
mainly in the senses oxygen need and
the physiologically
kidney by then triggers the synthesis
obligatory growth factor for
peritubular of EPO and its release into
erythroid development
cells the bloodstream

Through the interaction of EPO Thus EPO is a true

with receptor-bearing cells
within the bone marrow,
physiologic oxygen demands are
translated into increased red cell
production
hormone, manufactured at
one anatomic site and
transported through the
bloodstream to the site of
activity

EPO, in addition to promoting erythropoiesis directly, enhances

erythropoiesis indirectly by decreasing the interaction of
hematopoietic stem cells with their niches, reducing the amount of
trabecular bone and downregulating CXCR4 expression, the receptor
for CXCR12L/SDF1, on hematopoietic stem cell
 MICROENVIRONMENT OF
THE BONE MARROW

These are macrophages surrounded by

erythroid precursors in various stages of
development.
Hematopoiesis occurs in marrow
Since developing RBCs obtain iron via
cords, essentially a loose
transferrin, no direct contact with
arrangement of cells outside a
macrophages is needed for this.
dilated sinus area between the
arterioles that feed the bone and
Macrophages are now known to elaborate
the central vein that returns blood
cytokines that are vital to the maturation
to efferent veins.
process of the RBCs.
Erythropoiesis typically RBC precursors would not survive without
occurs in what are called macrophage support via such stimulation.
erythroid islands.
• A second role for macrophages in erythropoiesis also has been identified.
Although movement of cells through the marrow cords is sluggish,
developing cells would exit the marrow prematurely in the outflow were it
not for an anchoring system within the marrow that holds them there until
development is complete.

• There are three components to the anchoring system: a stable matrix of

accessory and stromal cells to which normoblasts can attach, bridging
(adhesive) molecules for that attachment, and receptors on the
erythrocyte membrane.

• The major cellular anchor for the RBCs is the macrophage. Several
systems of adhesive molecules and RBC receptors tie the developing RBCs
to the macrophages. At the same time, RBCs are anchored to the
extracellular matrix of the bone marrow, chiefly by fibronectin.
When it comes time for the RBCs to leave
the marrow, they cease production of the
receptors for the adhesive molecules.

Without the receptor, the cells are free to

move from the marrow into the venous
sinus.

Entering the venous sinus requires the RBC

to traverse the barrier created by the
adventitial cells on the cord side, the
basement membrane, and the endothelial
cells lining the sinus.

Egress through this barrier occurs between

adventitial cells, through holes
(fenestrations) in the basement membrane,
and through pores in the endothelial cells
Isla Eritroblástica: unidad anatómica
de la eritropoyesis; consiste en 2 Mf
rodeados de células eritroides en
maduración. La adhesión entre éstas
ocurre en estadio CFU-E y continua
durante toda la eritropoyesis.
En el desarrollo de la Isla eritroblástica,
participa la fibronectina,
hemaglutininas y sialo-adhesinas que
interactúan con receptors de Mf (EbR).

Conforme continua la maduración, los

precursores van abriéndose paso a través El núcleo es expulsado
de los macrófagos y, llegado el momento antes de su salida de la
de la expulsión del núcleo, el eritroblasto médula ósea, y es
se contacta con una célula endotelial, fagocitado y degradado
pasa a través de un poro en el citoplasma por los macrófagos.
de ésta e ingresa a la circulación.

La Isla Eritroblástica es una estructura de tal fragilidad, que

suele romperse en el proceso de aspiración de la médula ósea
con la aguja y, solo son visibles en muestras obtenidas por este
procedimiento cuando existe gran actividad eritriblástica
acelerada, como en las anemias hemolíticas y en la
eritroleucemia.
 ERYTHROCYTE
DESTRUCTION
• All cells experience the deterioration of their
enzymes over time due to natural catabolism.

• Most cells are able to replenish needed

enzymes and continue their cellular processes.

• As a nonnucleated cell, however, the mature

erythrocyte is unable to generate new proteins,
such as enzymes, so as its cellular functions
decline, the cell ultimately approaches death.

• The average RBC has sufficient enzyme function

to live 120 days.
• Because RBCs lack mitochondria, they
rely on anaerobic glycolysis for
production of ATP.

• The loss of glycolytic enzymes is

central to this process of cellular
aging, called senescence, which
culminates in phagocytosis by
macrophages, this is the major way in
which RBCs die normally.
• RBCs develop from committed erythroid progenitor cells in the bone marrow,
the BFU-E and CFU-E.

• The morphologically identifiable precursors of mature RBCs, in order from

youngest to oldest, are the pronormoblast, basophilic normoblast,
polychromatic normoblast, orthochromic normoblast, and polychromatic
erythrocyte or reticulocyte.

• As erythroid precursors age, the nucleus becomes condensed and ultimately

is ejected from the cell, which produces the polychromatic erythrocyte or
reticulocyte stage. The cytoplasm changes color from blue, reflecting
numerous ribosomes, to salmon-pink as hemoglobin accumulates and the
ribosomes are degraded. Each stage can be identified by the extent of these
nuclear and cytoplasmic changes.

• It takes approximately 18 to 21 days for the BFU-E to mature to an RBC, of

which about 6 days are spent as identifiable precursors in the bone marrow.
The mature erythrocyte has a life span of 120 days in the circulation.
• Hypoxia of peripheral blood is detected by the peritubular fibroblasts of the
kidney, which upregulates transcription of the EPO gene to increase the
production of EPO.

• EPO, the primary hormone that stimulates the production of erythrocytes,

is able to rescue the CFU-E from apoptosis, shorten the time between
mitoses of precursors, release reticulocytes from the marrow early, and
reduce the number of mitoses of precursors.

• Apoptosis is the mechanism by which an appropriate normal production

level of cells is controlled. Fas, the death receptor, is expressed by young
normoblasts, and FasL, the ligand, is expressed by older normoblasts. As long
as older cells mature slowly in the marrow, they induce the death of
unneeded younger cells.

• EPO rescues cells from apoptosis by stimulating the production of anti-

apoptotic molecules that counteract the effects of Fas and FasL and
simultaneously decreasing Fas production by young normoblasts.
• Survival of RBC precursors in the bone marrow depends on adhesive
molecules, such as fibronectin, and cytokines that are elaborated by
macrophages and other bone marrow stromal cells. RBCs are found in erythroid
islands, where erythroblasts at various stages of maturation surround a
macrophage.

• As RBC precursors mature, they lose adhesive molecule receptors and can
leave the bone marrow. Egress occurs betweenadventitial cells but through pores
in the endothelial cells of the venous sinus.

• Aged RBCs, or senescent cells, cannot regenerate catabolized enzymes because

they lack a nucleus. The semipermeable membrane becomes more permeable to
water, so the cell swells and becomes spherocytic and rigid. It becomes trapped
in the splenic sieve.

• Extravascular or macrophage-mediated hemolysis accounts for most normal

RBC death. The signals to macrophages that initiate RBC ingestion may include
binding of autologous IgG, expression of phosphatidylserine on the outer
membrane, cation balance changes, and CD47-thrombospondin 1 binding.

• Fragmentation or intravascular hemolysis results when mechanical factors

rupture the cell membrane while the cell is in the peripheral circulation. This
pathway accounts for a minor component of normal destruction of RBCs.