Hematopoiesis and Laboratory Hematology

Ted Strom, MD, PhD

Memphis VA Medical Center
UTHSC Dept of Pathology and Lab Medicine

Hematopoiesis: Differentiation and its

morphologies

Red cell production requirements

Laboratory evaluation of hematopoiesis

 Hematopoiesis:
Learning objectives:
Be able to correctly use the terms blast, pluripotent stem cell, burst
forming unit, and lineage-committed progenitor.

Be able to recognize the bone marrow cells responsible for production of

red cells, granulocytes, and platelets.

Be able to place, in the relevant growth and differentiation processes,

terms like: Megakaryocytes, promyelocytes, pronormoblast, band form.

Be familiar with the genetic and nutritional requirements for normal red
cell production, and how the process is regulated

Be able to recognize the key features (composition and cellularity) of a

bone marrow biopsy.
 Hematopoiesis and laboratory hematology

Ted Strom, MD, PhD

Memphis VA Medical Center
UTHSC Dept of Pathology and Lab Medicine

The complete blood count

Hematopoiesis: Differentiation and its

morphologies

Red cell production requirements

Laboratory evaluation of hematopoiesis

 Hematopoiesis:
Normal hematopoiesis:
In vivo differentiation processes

Regulated
Differentiation

and

Regulated
Cell division

~ 5 x 1011 cells/day
(excluding platelets)
 Hematopoiesis:
Normal hematopoiesis:
In vivo differentiation processes

Pluripotent stem cells

• Rare (<1 in 20 million)
• Can’t be clearly ID’d by morphology,
• Express receptors for key growth factors,
• Critical to bone marrow transplant and gene therapy
methods
 Hematopoiesis:
Normal hematopoiesis:
In vivo differentiation processes

Burst forming
units (BFU) and
colony forming
units (CFU):
currently defined
by their
responsiveness
to a handful of
key known
growth factors
 Hematopoiesis:
Normal hematopoiesis:
Key growth factors

Importance of
the regulated
differentiation/di
vision pathways:
We can
manipulate them
with drugs
mimicking the
circled growth
factors
 Hematopoiesis:
Normal hematopoiesis:
What we can see microscopically in the bone marrow

“blasts”
(< 4%)
 Hematopoiesis:
Normal morphologic maturation of granulocyte
precursors: (seen in aspirated material)

“Blast” (3-4%)

Promyelocyte (2%-8%)

Myelocyte (10- Metamyelocyte Bands and Neutrophils (25-40%)

13%) (10-15%)
 Hematopoiesis:
Normal maturation of red cell precursors: (seen
in aspirated material)

“Blast” (3-4%)

Pronormoblast

Basophilic Polychromatophilic Normochromic

erythroblast erythroblast erythroblast
 Hematopoiesis:
Normal maturation of erythroid precursors:
5 cell divisions

Mature
Red cells

Nascent
Red cells
- polychromasia
- reticulocytes

Extruded nuclei
 Hematopoiesis:
Megakaryocytes (thrombopoiesis): (usually seen
in “core” biopsy material)

Regulation:
“Blast” (3-4%)
Hepatocytes and
other sources

TPO

immature mature

Megakaryocytes
 Hematopoiesis:
Regulation of thrombopoiesis:
Thrombopoietin (TPO)

Arrows represent TPO “flux”

 Hematopoiesis and laboratory hematology

Ted Strom, MD, PhD

Memphis VA Medical Center
UTHSC Dept of Pathology and Lab Medicine

The complete blood count

Hematopoiesis: Differentiation and its

morphologies

Red cell production requirements

Laboratory evaluation of hematopoiesis

 Hematopoiesis:
Red cell production requirements

Requirements:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation

Heme synthesis requires:

The Porphyrin ring
Iron
 Hematopoiesis:
Red cell production requirements

Requirements:
Heme synthesis

Porphyrin
Iron
Globin synthesis
DNA synthesis
Regulation
Porphyrin synthesis requires:
Several enzymes:
Key players:
ALA synthetase
ferrochelatase
B6
Succinyl CoA
glycine
Which requires:
B12
Folate
 Hematopoiesis:
Red cell production requirements

Requirements:
Heme synthesis
Porphyrin
Iron
Globin synthesis
DNA synthesis
Regulation

Iron
Uptake system
Transport system
Storage system
Regulation of the above
Iron uptake, transport, and storage

Overview of Iron uptake and transport

Duodenal Reductase

dehydroascorbate Ascorbate

GI tract
(ferrous) iron Fe++ Dietary (ferric) iron Fe+++

plasma
(ferrous) iron Fe++ Fe+++ Fe+++/
transferrin

Multi-step process
Serum oxidases
Serum transferrin
 Anemia:
Iron uptake, transport, and storage Impaired red cell production

Iron transport and

storage Dietary iron

GI tract
Fe+++/transferrin

plasma
Transferrin receptor

Organs
Red cell precursors Storage pool iron (ferritin)
(bone marrow) Macrophages in liver, spleen, bone marrow
Iron uptake, transport, and storage

How iron uptake is

regulated

Dietary iron
(reduced; Fe++) DMT-1

GI tract
Duodenal enterocytes enterocyte
transferrin Serum oxidases

plasma
Ferroportin

Fe+++ Fe++

Fe+++/transferrin

Organs
Iron uptake, transport, and storage

Iron homeostasis:
liver

GI tract
(+) Fe+++/transferrin

plasma
Ferroportin

Hepcidin (+)

Organs
Storage pool
iron

internalization, degradation
 Hematopoiesis:
Red cell production requirements

Requirements:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation

Globin synthesis requires:

Normal globin genes
Alpha
Beta
Amino acids
 Hematopoiesis:
Red cell production requirements

Requirements:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation

DNA synthesis requires:

Deoxynucleoside triphosphates
Which require
Ribonucleotide reductase
Thymidine
Which requires:
B12
Folate
Red cell production requirements:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation

Three amino acids

(gln, glu, asp) Purines
(AMP, GMP)

PRPP

N10-formyl-THF

NADPH, CO2
Mechanisms for Impaired red cell production:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation
UMP TMP

N5, N10-methylene THF

Dihydrofolate
(DHF)

NADPH;
Dihydrofolate
reductase
Mechanisms for Impaired red cell production:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation

N5, N10-methylene THF

N5-methyl THF

homocysteine

Methionine
Mechanisms for Impaired red cell production:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation

N5-methyl THF

cysteine
homocysteine

Methionine
Red cell production requirements

Requirements:
Heme synthesis
Globin synthesis
DNA synthesis
Regulation

Erythropoietin:
Which requires:
Normal kidneys
A normal bone marrow micro-environment
 Hematopoiesis:
Type 1 hematopoietic growth factor receptors

Cytokine (Epo, Tpo)

Receptor (Epo-R; Tpo-R = c-MPL; GCSFR)

Cell membrane

Jak-2 kinase
 Hematopoiesis:
Type 1 hematopoietic growth factor receptors

℗ ℗ ℗ ℗

℗ ℗
 Hematopoiesis:
Type 1 hematopoietic growth factor receptors

℗ ℗ ℗ ℗ ℗℗

℗ ℗ ℗ ℗ ℗ ℗ ℗℗

STAT Phosphorylated Activated STAT

transcription STATs dimers
factors

Proliferation, Altered
differentiation transcription
patterns
 Hematopoiesis and laboratory hematology

Ted Strom, MD, PhD

Memphis VA Medical Center
UTHSC Dept of Pathology and Lab Medicine

The complete blood count

Hematopoiesis: Differentiation and its

morphologies

Red cell production requirements

Laboratory evaluation of hematopoiesis

 Anemia:
Iron uptake, transport, and storage Impaired red cell production

Lab
measurements Dietary iron

GI tract
Serum
Fe+++/transferrin
Iron

plasma
Transferrin receptor

Organs
Red cell precursors Storage pool iron (ferritin)
(bone marrow) Macrophages in liver, spleen, bone marrow
 Anemia:
Iron uptake, transport, and storage Impaired red cell production

Lab
measurements Dietary Fe+++

GI tract
Total iron
Fe+++/transferrin
binding

plasma
capacity
(TIBC)
transferrin

Transferrin receptor

Organs
Red cell precursors Storage pool iron (ferritin)
(bone marrow) Macrophages in liver, spleen, bone marrow
 Anemia:
Iron uptake, transport, and storage Impaired red cell production

Lab
measurements Dietary Fe+++

GI tract
Fe+++/transferrin

plasma
transferrin (serum ferritin)

Transferrin receptor

Organs
Red cell precursors Storage pool iron (ferritin)
(bone marrow) Macrophages in liver, spleen, bone marrow