RBC & Erythropoiesis

Dr. U. Rathna kumari MD.,DNB

Hematocrit (PCV) tube
 Blood Cells
Blood cells are erythrocyte (red blood cell, RBC), leukocyte (white
blood cell, WBC) and thrombocyte (platelet, P).
 Blood Cells
• The forming processes of erythrocyte (red blood
cell, RBC), leukocyte (white blood cell, WBC) and
thrombocyte (platelet, P) originating from
hematopoietic stem cells are hemopoiesis.
 Hemopoietic process
• yolk sac hemopoiesis (early embryo period)
• liver and spleen (second embryo month)
• marrow↑ and liver, spleen↓ (after fourth
embryo month)
• marrow (fetus birth time) and liver, spleen as
complementary role.
During adulthood (after 18), red marrow (flat
bones, e.g. vertebra,ilium, sternum, rib, skull
and long bone ending)
 Hemopoietic process
Stage one: Hemopoietic stem cells
self renewal, steady numbers, active differentiation.

Stage two: committed progenitors, directional

differentiation (CFU-GEMM, CFU-E, CFU-GM, CFU-MK,
CFU-TB). [CFU: colony- forming unit]
Stage three: precursors
morphologic occurrence of various original blood cells.
 Hemopoietic stem cells
Basic characteristics
• Self renewal in high degree, constant from young to old age.
• Multi- directional differentiation
• Large potential proliferation, Hemopoietic stem cells produce
about 1×1011 blood cells releasing to blood for use.
• Surface sign
According to CFU (colony forming unit), using fluorescence-
activated cell sorting (FACS), its main surface sign is CD34+CD38-
Lin-and CD34-CD38-Lin-.
Note
CD: cluster of differentiation of antigen on the white blood cells;
Lin: systemic specific antigen on the hemopoietic cells.
 Hemopoietic microenvironment

• It includes stromal cell secreting extracellular

matrix (ECM), multihemopoietic regulating
factor, hemopoietic nerves and blood vessels.
• Stromal cells in the marrow come from fibrocyte,
reticulocyte, endothelial cell, ectoblast cell,
monocyte, engulfing cell, osteoblast and
osteoclast.
• Stromal cells supply two material: one is soluble
hemopoietic growth factor, another is
membrane-combined adhesive molecule.
• Extracellular stroma synthesized and
secreted by marrow stromal cell filling
cellular interstice contains big molecules,
such as collagen (typeI, II, III, IV),
glycoprotein (fibronectin, laminin,
hemopoieticnectin ) and protein amylose
(sulfate cartilagetin, sulfate heparin,
hyaluronic acid and sulfate dermatin, etc).
• Hemopoietic cells must adhere to stromal
cell and is in the hemopoietic
microenvironment for survival.
Hemopoietic process
Hemopoietic process
 Erythrocyte Physiology
Shape and number of red blood cells (RBC)
• Shape of RBC: like biconcave disc

Its diameter is about 7~8 µm, peripheral thickness about

2.5 µm, central thickness about 1 µm and cubage about
90 µm3.
 Reason for shape of RBC

biconcave disc like

 Erythrocyte Physiology
Number of RBC: It is most numbers in the blood.
Male adult, 4.5~5.5×1012/L; average, 5.0×1012/L
Female adult, 3.8~4.6× 1012/L; average, 4.2×1012/L
Newborn, ≥ 6.0×1012/L
Pregnant female, numbers of RBC and Hb are relatively less
(because of more plasma).
Dweller in hills, numbers of RBC and Hb are relatively more
(because of compensation for anoxia).
 Physiological Characteristics and
Functions of RBC
① Permeability: semipermeable membrane, gas and urea
freely passing through, negative ions easily in or out of
RBC, and positive ions not. There are Na-K ATPase as
pump on the membrane of RBC
② Plasticity and metamorphose:

Plasticity and metamorphose depend on: 1) surface area-cubage ratio, 2)

viscosity of Hb, 3) membrane elasticity and viscosity.
Physiological Characteristics and
Functions of RBC
Functions of RBC
• RBC can be used for transportation of
O2 and CO2 in the blood.
• RBC can be served as pH buffer.
 Life and breakage of RBC
• Life-span: 120 days, about 4 months, each RBC circulates 27 km
averagely in vessels, short life-span for aged RBC
• Breakage: places are liver, spleen and lymphatic node, and after
breakage, Hb released from RBC immediately combine with
plasma α2-globulin (Hb touched protein) which is taken in by
liver for iron reuse.
• Hb, very toxic if it get into blood, normally, it can be metabolized
into bile pigment in liver.
• Clinic relation.
 Hemopoiesis

• the process of production of all formed elements by

the bone marrow
Stem cell:
Bone marrow has very specialized type of cells called
stem cell. They have the
• remarkable ability to produce many types of other
cell of the body.
• When stem cell divides, it has the potential to
remain as a stem cell or turn into another type of
cell with differentiated and specialized functions.
The process of formation of
RBCs is called Erythropoiesis
 RBC Formation before birth
• Mesoblastic stage
– Nucleated RBCs - Yolk sac and
Mesothelial layers of the placenta –
3rd week
• Hepatic stage
• At 6 weeks - Liver form blood cells
– Spleen + lymphoid tissues form blood
cells.
 RBC Formation before birth

• Myeloid stage
• From the third month onwards - the bone

marrow gradually becomes the principal

source of the RBCs

• Last month – Bone marrow exclusively

 RBC Formation after birth
• The bone marrow - all bones - 5 years
• Marrow of the long bones (except for the
proximal humerus and tibia)

• Most red cells continue to be produced in the

marrow of the membranous/ flat bones, such
as
– Vertebrae, Sternum, Ribs, and Ilium.
Relative rates of red blood cell production in the bone marrow
of different bones at different ages.
 Bone marrow cells for Erythropoiesis

• Pluripotential hematopoietic stem cell, PHSC

• Committed stem cell that produces erythrocytes is
called
Colony-forming unit–erythrocyte, CFU-E

Factors:
– Growth inducers
– Differentiation inducers.
 ERYTHROPOIESIS
PHSC
Bone marrow

CFU-E 4-5 days

Proerythroblast
Polychromatophil erythroblast
Orthochromatophil erythroblast

Reticulocyte
Blood 1-2 days.
Erythrocyte.
 Proerythroblast

• No hemoglobin

• Nucleus 12 um

• Contain nucleoli
 Basophil erythroblast
• Early normoblast
• Nucleoli disappear
• Show mitosis
• Cytoplasm deep blue
– Increase in RNA

• Hemoglobin starts
appearing – Little Hb
 Polychromatophil erythroblast
• Late normoblast

• Nucleus smaller

• Coarse Chromatin

• Hemoglobin increase
– Eosinophil Stain

• RNA – Basophil stain

 Orthochromatic Erythroblast

• Normoblast

• Nucleus smaller

– Pyknosis

• Nuclear lysis and

• Nuclear extrusion
 Reticulocyte
• Reticulum
• Remnant of ER & GA
– Synthesize Hb

• Few Mitochondria
• Young RBCs (34% Hb)
• 1 % of Red Cells
• Importance?
 Requirements of Erythropoiesis

• Erythropoietin.
Availability of other specific nutrients.
• Vitamins e.g. vitamin B12, vitamin B6, Thiamine,
• Riboflavin, Vitamin C and E.
• Metals :
• Iron for hemoglobin synthesis
• Trace metal such as cobalt
• Proteins particularly the essential amino acids
 Transfer of RBC to Circulation
RBC pass from the bone marrow into the blood
capillaries

By

Diapedesis
(squeezing through the pores of the
capillary membrane).
 Erythrocytes

• Round, biconcave, disc shaped.

• Smooth contours
• Diameter 7.8 um.
• Normally no variation in size
and shape.
• Stain with EOSIN.
– More stain at periphery
• Can deform easily.
 STRUCTURE OF RBC.
• Negative surface charge.
• Bag of fluid with dissolved substances and
hemoglobin
• Membrane –
– Outer glycoprotein coat
– Lipid bilayer (PL 55%,Cholesterol 45%)
• Inner protein molecules cytoskeleton
– Spectrin, Actin, Ankyrin etc.
• No sub cellular particles
 ENERGY METABOLISM

• Less energy required

– Na + - K + pump
– Iron in Fe ++ form

• Utilize Glucose by GLUT 1

• Glycolysis
– Embden Meyerhof pathway

• Pentose phosphate pathway.

– Hexose monophosphate shunt
 Features of a Mature RBC
• Biconcave disc
• Mean Diameter 7.4 micron
• Can deform easily.
• Bag of fluid with dissolved substances and
hemoglobin
• No sub cellular particles
• Metabolism
– Anaerobic respiration- Glycolysis
– Pentose phosphate pathway.