https://www.youtube.com/watch?

v=oTsGjsOcNPQ
Hematopoiesis
(blood cell formation)
 Hematopoiesis: A quick review

Learning objectives:
- Get to know what is Hematopoiesis
- Understand where, when, & why does it take place
- Get acquainted with stem cells especially as they relate to hematopoiesis
- Become familiar with growth factors & interleukins involved in
hematopoiesis
 Why do we need the Hematopoiesis?

– Cells of the blood are constantly being lost or

destroyed. Thus, to maintain homeostasis, the
system must have the capacity for self renewal.
Kinds of Stem Cells
Stem cell
type Description Examples

Cells from early (1-3

Totipotent -give rise to a whole organism
days) embryos

Some cells of
-give rise to the 3 major body
Pluripotent layers (ecto, meso & endoderm blastocyst (5 to 14
days)

-(hematopoietic and others): Fetal tissue, cord

Multipotent  give rise to a limited blood, and adult stem
number of cell types cells
Stem cells
Multipotent (hematopoietic) Stem Cells

• Undifferentiated and uncommitted stem cells residing mostly

in the bone marrow
• Have the ability to self-renew or differentiate into any of the
many blood cell types. 
• When stimulated with the proper stimulus, such cells will
differentiate into the myeloid lineage or lymphoid lineage.
Sites of production:

• Developing cells situated outside of BM sinuses  mature cells

released into sinus spaces  marrow microcirculation  general
circulation.
 Yolk Sac: a store of
nourishment for the
Early life
 embryo

Later on

Localization of hematopoiesis to bone
marrow
Bone Marrow Stroma
• Suitable environment for
Stem Cell (SC) growth & def.
• Composed of stromal cells +
microvascular network.

Extracellular molecules:
Stromal cells: • Collagen
• adipocytes • Glycoprotein (fibronectin,
• Fibroblast thrombospondin)
secrete
• Reticulum cells • Glycosaminoglycans
• Endothelial cells (hyaluronic acid &
chondroitin derivates)
• Macrophages • Growth factors  for cell survival
Hemopoietic Growth Factors (HGF)
• Glycoprotein hormones  regulate proliferation & differentiation of
HPC & function of mature blood cells.
• Biological effects of HGF mediated through specific receptors on
target cells.
• Activity:
– Locally  at the site where they are produce  by cell-cell contact.
– Circulate in plasma
HGF
• May bind to EC matrix  form niches to which SC adhere.
• Major sources (except erythropoietin):
• T-lymphocytes
• Monocytes (& macrophages)
• Stromal cells
• Erythropoietin  90% synthesized by peritubular capillary lining cells
within the kidney
• Other erythropoietin is synthesized by hepatocytes
• Thrombopoietin  largely made in liver
Hematopoiesis
microenvironment(s)
Growth factors:
CSF, colony‐stimulating factor; FLT3‐
L, FLT3 ligand; G‐CSF, granulocyte
colony‐stimulating factor; GM‐CSF,
granulocyte–macrophage colony‐
stimulating factor; IL, interleukin;
M‐CSF, macrophage colony‐
stimulating factor; SCF, stem cell
factor; TNF, tumor necrosis factor;
VEGF, vascular endothelial growth
factor. * These also act
synergistically with early acting
factors on pluripotential
progenitors.
Figure 1.6 A diagram of the role of growth factors in normal hemopoiesis. Multiple growth factors act
on the earlier marrow stem and progenitor cells. EPO, erythropoietin; PSC, pluripotential stem cell;
SCF, stem cell factor; TPO, thrombopoietin; FLT3‐L, FLT3 ligand. For other abbreviations see Fig. 1.2.
Granulocyte-macrophage colony-stimulating factor (GM-CSF)
fms-like tyrosine kinase 3 (FLt3) receptor
Function of
Growth Factors:

Figure 1.5 Growth factors may

stimulate proliferation of early
bone marrow cells, direct
differentiation to one or other cell
type, stimulate cell maturation,
suppress apoptosis or affect the
function of mature non‐dividing
cells, as illustrated here for
granulocyte colony‐stimulating
factor (G‐CSF) for an early myeloid
progenitor and a neutrophil
Adhesion Molecules
• Glycoprotein
• Mediate the attachment of marrow precursors, leukocytes and
platelet to various components of the extracellular matrix :
• Endothelium
• Other surfaces
• Each other
– Hematopoiesis in the bone marrow is called medullary
hematopoiesis
– Hematopoiesis in areas other then the bone marrow is called
extramedullary hematopoiesis
– Extramedullary hematopoiesis may occur in fetal hematopoietic
tissue (liver and spleen), and in adult when the bone marrow cannot
meet the needs of the tissues. This can lead to hepatomegaly
and/or splenomegaly (increase in size of the liver or spleen because
of increased functions in the organs).
 Types of hematopoiesis
Myelopoiesis:
• Erythropoiesis: production of RBCs; 5-38% of nucleated cells in BM.
• Megakaryopiesis: occurs adjacent to sinus endothelium (blood vasculature
inside the bone tissue); megakaryocytes protrude through the vascular wall as
small cytoplasmic processes to deliver platelets into blood, Megakaryocytes
develop into platelets in approximately 5 days.
• Granulopoiesis: production of neutrophils, eosinophils, & basophils in BM;
maturing cells stay for ~ 6 days in the proliferating pool. 
Lymphopoiesis:
1-5% of nucleated cells are lymphocytes and plasma cells; produced in lymphoid
follicles within the BM.
Normal Cell Maturation
1) Changes in cytoplasm
2) Changes in nucleus
3) Reduction in cell size
4) granulations

Gradual transformation
Changes are simultaneous and parallel
General cellular characteristics of maturing blood cells
1. Cell size:
•Overall size of all blood cells decrease with maturation except for that of
megakaryotes
•Hence, presence of larger than normal cells in blood may indicate presence of
immature cells  further investigation.
2. Nuclear-cytoplasmic (N:C) ratio:
•Defined as amount of nuclear space to that of cytoplasmic space.
•As size of nucleus generally decreases with maturation so does the N:C ratio.
•The N:C ratio for blast forms of erythrocytes, leukocytes, and megakaryocytes
is 4:1 but as these cells mature, the ratio drops to 2:1 or even 1:1.
•Note that mature RBCs and thrombocytes are anuclear  all are cytoplasm;
•mature lymphocytes tend to maintain larger than normal N:C (3-4:1).
3.Nuclear Characteristics

1. Chromatin pattern:

As blood cells mature, chromatin

changes from loose looking
arrangement to a more dense,
clumped or pykontic (compact)
pattern.
2. Nuclear shape:
In young cells nucleus is either round or oval; monocytes may have a slightly folded
nucleus. For cells that retain the nucleus, its shape becomes distinct for each specific
cell (e.g. round/oval for lymphocytes, segmented for neutrophils, etc.)
3. Presence of nucleoli:

Erythrocytes, leukocytes and megakaryocytes have nucleoli in the earliest

recognizable stages of blood cell development.
1&3 / nucleus may be apparent, they disappears in mature cells.
 4.Cytoplasmic Characteristics
Staining Characteristics of blood cells:
Wright Stain: Methylene blue (basic component) and Eosin (acidic
component)
Basic cellular elements react with acidic component (eosin):
Acidophilic
Acidic cellular elements react with basic component (methylene
blue): Basophilic
Neutral elements of cell react with both components: Neutrophilic
 Nuclear DNA and cytoplasmic RNA are acidic (basophilic): Stain blue
 Hemoglobin in red cells is basic (acidophilic): Stains orange red
 Proteins and enzymes in granules vary  overall color varies depending
on the % of acidic vs. basic constituents.
• Cytoplasm components..

• RNA content, stages  ???

• Protein content  ?

• Basophilic

• Eosinophilic

• Heamoglobin is ____philic
Source: http://slideplayer.com/slide/4146928/
 Cytoplasmic Characteristics
1. Staining color and intensity: In Wright-stained blood films, color of
cytoplasm changes from darker blue (high nucleic acid [& protein]
content) in immature cells to lighter blue, blue-gray, or pink. Immature
erythrocytes have a very distinctive dark blue cytoplasm that then
changes to gray as the cell starts to make Hb and then turns pink in
mature RBCs.

2. Granulation: Immature cells have no granules, but nonspecific granules

of various shapes and colors start to appear as cells mature  each type
of mature cells exhibit a number of specific types of granules.
Red/burgundy granules: azurophilic; Blue: Basophilic; Orange: eosinophilic

3. Vacuoles: Number of vacuoles Increases as the cell ages as in the case

of Monocytes contains vacuoles.