HEMATOPOIESIS PART I
Hematopoiesis
- Blood production
Functions of the blood
- Carry and transport nutrients for
cells to produce energy
- Hemostasis
o Through blood clotting
- Healing of wounds
o Protein coagulation
o Presence of platelets
- Gas transport
o Oxygen and Carbon dioxide
(product of metabolism) for
cell metabolism
- Fight against pathogenic organisms
o Through white blood cells
▪ Innate immunity
▪ Adaptive immunity
- Buffering of the blood
o Maintain the normal pH
o Normal blood pH: 7.35-7.45
▪ Below 7.35 = acidosis
General characteristics of the blood:
- Average: 5-6 L
o 5L – female
o 6L – male
- 7-8% of the body weight
- 1 gram of Hemoglobin carries
1.34ml of Oxygen
o 600g of hemoglobin is
present in the entire body of
an adult
▪ Can carry around
800ml of Oxygen
- 45% formed elements
- 55% fluid portion
o Plasma – uncoagulated
blood
o Serum – coagulated blood
- Slightly alkaline
Plasma vs. Serum
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Plasma
- Has clotting factors
o Fibrinogen groups
o Protein
- Present clotting factors:
o 1,5, 8, 13
o These are absent in serum
Hematopoiesis
- General term for blood production
- Erythropoiesis – RBC production
- Leukopoiesis – WBC production
- Megakaryopoiesis – platelet
production
3 periods of hematopoiesis
1. Mesoblastic period
a. First trimester of pregnancy
(embryonic life)
2. Hepatic period
3. Myeloid period
* the first two phases, blood is produced
outside the bone marrow (extramedullary
hematopoiesis)
* if bone marrow is the one producing the
blood cells (medullary period of
hematopoiesis)
Detailed description of each period
Mesoblastic period
• Start of production: 19th – 20th day of
pregnancy
o First trimester of pregnancy
• Source: yolk sac (main)
• Kind of blood produced: only RBCs
o Embryonic / Primitive RBCs
▪ Do not have the
capability to remove
its nucleus
• Type of Hemoglobin: 3 types
(embryonic Hgb)
HEMATOPOIESIS PART I GKLDB 3MT4

o Portland o These organs are important

o Gower 1 to the maturation of
o Gower 2 lymphocytes
• Globin chains present: − Kind of blood produced: RBCs &
o Portland – 2 zeta & 2 gamma other blood cells
o Gower 1 – 2 zeta & 2 epsilon o WBCs
o Gower 2 - 2 alpha & 2 o Platelets
epsilon − Type of Hemoglobin: Hgb F (fetal
• End of production: before the 3rd Hgb)
month of pregnancy − Globin chains present: Hgb F
o 2 alpha & 2 gamma
Hemoglobin
− End of production: until 1st – 2nd
- Protein with 2 entities week after delivery
o Heme
Other details:
o Globin – protein part of Hgb
− Made up of amino acids − Hemoglobin F
− Can assume quaternary structure o Concentration: may reach up
o With bends to 60 – 90%
− Any kind of Hgb always has 4 globin o BUT for newborns, Hgb F is
chains: expected to be present
o α - alpha ONLY IN SMALL
o β – beta CONCENTRATION
o γ - gamma o Hypoxia – if adults still has a
o ε – epsilon HIGH AMOUNT OF HGB F
o δ – delta ▪ They appear cyanotic
o ζ – zeta (bluish)

Hepatic period of hematopoiesis Why should Hgb F be removed in adults (or

if present, in small amount only)?
− Liver that assumes the responsibility
to produce blood − Hgb F is not effective in
− Start of production: around 5th – 6th transporting Oxygen in the body
week of pregnancy tissue
o Peak of production: 5th – 6th − Hgb F holds Oxygen more instead
month of pregnancy of releasing it
− Source: Liver (main) o Leads to low oxygenation
− Other organs that support blood dev.
Disorders related to an increase in Hgb F:
(WBCs development)
o Spleen − Thalassemia
o Lymph nodes o Quantitative Hgb defect
o Thymus wherein a globin chain is
▪ Important for the reduced or is completely lost
differentiation of o Insufficient production of the
lymphocytes globin chains
HEMATOPOIESIS PART I GKLDB 3MT4

▪ No or less synthesis o Hgb A2- 2 alpha & 2 delta

o Minor thalassemia – reduced − End of production: death
number of globin chains
Other information:
o Major thalassemia – no
globin chain at all - Red bone marrow
− Example: Beta thalassemia major o Distributed in different body
o Beta globin chain is totally parts – in newborns
lost o Long bones in newborns are
o Affects the hemoglobin types replaced by the yellow
with beta in its composition marrow
▪ Red BM can only be
seen in the distal
Hereditary persistent fetal hemoglobin
portions of the long
− There is an increase in Hgb F in bones
adults o In adults: red BM is present
− Hgb F should not be present in in flat bones (sternum, pelvis,
adults anymore scapula)
− Adults patients diagnosed with this: - Thalassemia:
100% Hgb F o Alpha thalassemia
o Instead of A1 and A2 ▪ Affects A1, A2 & F
▪ Major: affected globin
Hgb switching – hemoglobin type changes chains will be
for every source replaced by abnormal
Myeloid period/ Medullary period of globin
hemopoiesis o Beta thalassemia
▪ Affects A1 levels
− Start of production: 4th – 5th month of o The effect of thalassemia
pregnancy depends whether it’s minor
− Source: Bone marrow or major
− Kind of blood produced: all types of - Decrease in Hgb A1 concentration
blood cells leads to anemia or cyanosis
− Types of Hemoglobin: (A- adult)
o Hgb A1
▪ Concentration: 97-
98%
▪ Most effective Hgb in
the adult body
o Hgb A2
▪ Concentration: 2-3%
− It is expected that adults should not
have Hgb F anymore (or it should be
less than 1%)
− Globin chains present:
o Hgb A1 – 2 alpha & 2 beta
HEMATOPOIESIS PART I GKLDB 3MT4

o Hepatic: Hgb F
o Myeloid: Hgb A1 and A2
- Its concentration is increasing
- Even in adult life, it is still present

C – gamma (γ)

- After hepatic, its production is

decreased

D – beta (β)

- Production starts to increase in

myeloid phase
- Hemoglobin A1
- Increasing concentration, almost the
same as alpha during adult life

E – delta (δ)

- Present in small amounts during

myeloid phase – A2

Summary of globin chains

Molecular
Hemoglobin Stage of life
Structure
Portland ζ2γ2 Embryonic
Gower I ζ2ε2 Embryonic
Gower II α2ε2 Embryonic
Newborn &
Hgb F α2γ2
adult
Newborn &
Hgb A1 α2β2
adult
Newborn &
Hgb A2 α2δ2
adult

Flow of globin chain production: Theories on blood production

A – zeta (ζ) & epsilon (ε) Monophyletic theory
- Their concentration is decreasing - All blood cells come from one single
- Only present in mesoblastic phase cell
B – alpha (α) o Multipotent hematopoietic
stem cell
- As early as fetal life, it is already ▪ There is a lot
present potential that this can
o Mesoblastic: Gower 2 become other cells
HEMATOPOIESIS PART I
- Most acceptable theory
Dualistic theory
- Cells originate from 2 stem cells
Poylphyletic theory
- Three or more stem cells which are
separated from each other
o Myeloid stem cell
o Erythroid stem cell
o Lymphoid stem cell
Hematopoietic stem cells
- If the stem cells produce blood
- Have unique CD markers
o CD – cluster of designation/
differentiation
o CD can be identified by flow
cytometrical analysis
o Used to identify a specific
cell
o Example: type of lymphocyte
with CD4+ - T-helper
lymphocyte
▪ HIV – attacks the T-
helper cell; lab test
assessment: CD4
count (# of T-helper in
the blood)
− Stem cell markers:
o CD 34
o CD 33
o HLADR
o Molecules that pertain to
blood stem cells
o Blood cells are immature
▪ On its first stage
o During maturation, these
markers are lost/ replaced
- CD8 – T-cytotoxic cells
3 fates of the hematopoietic stem
cells:
1. For renewal
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2. For differentiation
3. For apoptosis – natural cell death
− If the stem cell divide, it produces 2
daughter cells
− Renewal: If the 2 daughter cells are
reserved into the stem cell pool
▪ Reserved for renewal
− Differentiation: if the 2 daughter
cells differentiate/ divide into
different formed elements
o Symmetric Hematopoiesis /
division – if the 2 daughter
cells divide at the same time
o Asymmetric
Hematopoiesis – if only one
daughter cell differentiates,
the other either undergoes
apoptosis or renewal
▪ if the fate of the 2
daughter cells differs
− Apoptosis: if the 2 daughter cells
died, when cells self-destruct
(autolysis)
o Different from necrosis – cell
death due to trauma/ injury
o Apoptosis is a natural
physiologic way to remove
excess cells in the body
▪ Increase in cells =
overproduction of
cells
− Polycythemia vera
o Thick blood
o Bone marrow keeps on
producing formed elements
even if unnecessary
o Increase number of elements
than necessary
o Clog blood vessels
Theory (what influences the cells’ fate?)
1. Stochastic model of hematopoiesis
HEMATOPOIESIS PART I GKLDB 3MT4

a. Random – depends on the o Early acting

cells whether they will die/ o Intermediate acting
differentiate o Late acting
2. Instructive model of hematopoiesis − Early acting
a. Dictated by the o Commonly activating the
microenvironmental earliest sign of blood cell
factors within the bone development
marrow o SF – stem cell factor/ kit
b. There are factors that ligand
influences the stem cells on o FLT3 ligand
their fates
Intrinsic factors
c. Lineage differentiation/
Multilineage differentiation − Cells carry genes that regulate their
differentiation/ apoptosis
− GATA 2 gene
In the instructive model: − TAL 1 gene (TL1)
o Genes within the cells that
− Stem cells receive signals
can progress/ repress the
− 2 types of signals:
development of the cell
o Extrinsic signals –
influencers that are outside Genes that can regulate the stem cells:
the stem cell (additional genes)
o Intrinsic signals – factors
within the cells (regulatory − Notch 1 gene
genes) − Notch 2 gene
− Extrinsic signals Stem cells because of the bone marrow
o Cell to cell – cell near the influences, it can become myeloid or
bone marrow may cross-link, lymphoid stem cells
may affect one cell
o Growth factors − Depends on the growth factors that
− GF: If present, it will influence the are combined in the bone marrow
cell to differentiate to a specific type
of cell
o Cytokines
o IL (interleukin)
▪ One kind of cytokine
(chemicals released
by a cell)
o CSF – colony stimulating
factor
o SF – stem cell factor
− 3 kinds of growth factors
(necessary for the maturation of the
blood cells)
HEMATOPOIESIS PART I GKLDB 3MT4

o Myeloid – RBC, WBC,

platelets
o Lymphoid – T, B, or NK
lymphocyte

But if the cells will produce specific type of

formed elements, they are now committed

− Cells will not differentiate into other

cells beside the specific type of cell

For platelet production (additional from

other books): IL-11 & IL-7

− TPO – thrombopoietin (specific

cytokine needed)
− Lymphoid stem cell – mature = For RBC: Epo – erythropoietin
lymphocytes
o B-cells − Epo is needed for the myeloid to be
o T-cells committed to RBCs
o NK-cells For WBC: GM-CSF: granulocyte monocyte
− Myeloid stem cell, it can transform to colony stimulating factor
different blood cells (almost all
formed elements) − Myeloblast – 1st stage of WBCs
o Platelets production
GF needed for differentiation of basophil,
o RBC production
neutrophil, monocytes: IL-3 & IL-6
o WBC production
− Leukemia – classified as to where GF for differentiation of eosinophil: IL-3 &
the cell comes from IL-5
o Lymphoma – cancer of the
lymphoma
▪ Classification: 5 stages of mitosis
lymphoproliferative
G0 – resting stage of cells
o Erythroleukemia – immature
RBCs G1 – when cells are activated to divide
▪ Myeloproliferative –
erythro comes from − Cells are gathering the information,
myeloid stem cell materials for them to divide

Multipotent hematopoietic stem cell – still S phase – DNA replication is present

uncommitted because it can differentiate G2 – cells are ready to divide
into myeloid or lymphoid
M phase – cells are dividing (mitosis)
− Lymphoid and myeloid stem
cells are also uncommitted
IL 3 – myeloid progenitor stimulating factor
HEMATOPOIESIS PART I
− In the myeloid, from the beginning
IL-3 is present
− Progenitor – immature cells
− IL-3 is derived from T-cells
Erythropoiesis
- Red blood cell production
- Stimulated by the erythropoietin Epo
o Released solely in the kidney
▪ Kidney failure –
possible anemia
▪ Kidney – main source
of Epo
- Functions of the Epo:
o Development of RBCs
o Early release of RBCs from
the bone marrow to the
circulation
o Preventing apoptosis (cell
death)
o Reducing the length of time
for cell production
- General length of time (from
rubriblast to mature RBC): 5 days
(3-6 days)
o Generally, it takes one week
o In some cases, time gets
shorter when the body needs
it (or when you are
experiencing hypoxia)
o Epo will stimulate the bone
marrow to release RBCs to
the circulation reducing the
time of maturation
▪ Days depend on the
severity of the
disease
▪ Like thalassemia
major
- 2 early stage of RBC (before
rubriblast):
o BFUE – burst forming unit
erythroblast
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o CFEU – colony forming unit
erythroblast
- Considering the stage from BFUE to
mature RBC: it takes 18-21 days
o Depending on the influence
of Epo
- From rubriblast to mature RBC: 4
mitotic divisions (average)
Rubriblast – earliest stage you can identify
using the light microscope
- BFUE & CFUE – hard to distinguish/
differentiate from the myeloid stem
cells
Nomenclature types:
- Rubri nomenclature
o Using “rubri” in naming
▪ Rubri = red
- Ehrlich nomenclature
o Uses “erythro”
- Normo nomenclature
o Uses “normo”
Names: (6 stages – from rubriblast)
- Rubriblast
o Proerythroblast
o Pronormoblast
- Prorubricyte
o Basophilic erythroblast
o Basophilic erythroblast
- Rubricyte
o Polychromatophilic
erythroblast
o Polychromatophilic
normoblast
- Metarubricyte
o Orthophilic erythroblast
o Orthophilic normoblast
- Reticulocyte
o Diffusely basophilic
erythrocyte (or normocyte)
o Polychromatophilic
erythrocyte (normocyte)
HEMATOPOIESIS PART I GKLDB 3MT4

o Has no nucleus = X “blast” - Nucleoli is not visible (but still

- Mature red blood cells present)
o Erythrocyte - NC ratio: 6:1
o Normocyte o Cells size is decreasing
o Nucleus is decreasing
Characteristics of each stage
- Average size: 15-17 um
Rubriblast
Question: in what stage euchromatin &
- Cytoplasmic color: blue (basophilic) parachromatin is visible? As early as
- Chromatin material: fine and lazy where? prorubricyte
o Evenly distributed all
- Stays in the bone marrow for 24
throughout the nucleoplasm
hours
- Has visible nucleoli
- GF is continuous for cell
- Average size: 17-25 um
development
- NC ratio: 8:1
- Prorubricyte will divide twice
o Nuclear cytoplasmic ratio
o 1 prorubricyte = 4 rubricyte
o Increased – because the
- So, the 2 prorubicytes produced by
nucleus is almost occupying
the rubriblast will produce 8
the entire diameter of the cell
rubricytes
- Stays in the bone marrow for 24
hours Rubricyte
- Number of mitotic division (from
- Polychromatophilic erythroblast
rubriblast to prorubricyte): 1
o Cytoplasm tries to become
o Produce 2 prorubricyte
red (reddish appearance)
daughter cells
- Cytoplasmic color: red-blue (purple)/
Prorubricyte reddish
o 2 shades of color
- Cytoplasmic color: blue (basophilic)
o Hemoglobin is already in the
- Chromatin material: slight
cytoplasm of the rubricyte
compactness
▪ Hgb gives the
o Chromatin material is trying
redness color
to compact
o Hemoglobin is already whole
o This is what is happening in
in the cytoplasm
karyorepsis
- Nucleus is not yet dead
▪ Nucleus degeneration
o Nucleus is still shrinking
o There are white & dark areas
- Chromatin material: moderate
(stained & unstained)
compactness
▪ Presence of
o Continuous degeneration of
euchromatin &
the nucleus
parachromatin
- NC ratio: 4:1
o But in here the chromatin
- Divides once to metarubricyte
material is still clumping
o Produces 16 metarubricyte
(nucleus is not yet
- Last stage where division happens
degenerated)
HEMATOPOIESIS PART I GKLDB 3MT4

o Because karyorepsis is o If nucleus is already

already present in the exposed, it will release “eat
metarubricyte me” molecules
▪ Phosphatidyl serine
Hemoglobin synthesis starts where?
• Phagocytic
Rubriblast
factor
Hemoglobin is totally complete in what ▪ Molecules will be
stage? Rubricyte recognized by the
macrophages in the
Metarubricyte
bone marrow
- Stage where cells will not divide ▪ Nucleus will be
anymore (no nuclear division) removed & eaten
o Nucleus is already o Phosphatidyl serine – can
degenerated dictate the macrophage to
- Cytoplasmic color: has shades of eat the nucleus
grayish, reddish, and bluish ▪ WBCs need
appearance stimulating factors
o Orthochromatophilic o Pyrenocyte – RBC with
- Chromatin material: heavy exposed nucleus
compactness (highly compact) ▪ Metarubricyte na
- NC ratio: 1:2 pinipinch off yung
o Dead nucleus nucleus
o Reverse NC ▪ Metarubricyte with
- Average size: 8-12 um extruding out nucleus
▪ Phagocytes will eat
Reticulocyte the nucleus
- Has no nucleus - In certain disorder, sometimes the
pinching of nucleus is not complete
As the red cell develops, the average size & o Remnants of nucleus =
NC ratio is decreasing Howell-jolly bodies
What causes the removal of the nucleus?

- Vimentin – special component that

holds the organelles of the maturing
RBCs
- As the RBCs mature, because of the
Epo stimulation, Vimentin is lost
o Results to loosening of the
organelles in the maturing
cells
o Lumalabas yung nucleus
o ‘Pinching’ will occur =
removal