Module 4: RBC
Maturation and Function
Acknowledgments
Ministry of Health – Guyana
Centers for Disease Control and Prevention
(CDC) / Global AIDS Program (GAP) – Guyana
Centers for Disease Control and Prevention
(CDC) – Atlanta
American Society for Clinical Pathology (ASCP)
Objectives
Review the origin of blood cells using the present stem
theory of haematopoiesis.
Describe factors involved in the regulation of erythropoietic
activity, including substances needed for erythropoiesis and
haemoglobin synthesis.
Discuss the site of synthesis of erythropoietin and its
influence on erythropoiesis.
List two nomenclatures used to describe the stages of RBC
maturation.
Objectives
Differentiate each cell in the RBC maturation series based on
cell size, nuclear size, cytoplasmic color and nuclear
chromatin.
Describe the reticulocyte, including its composition, time
spent in the bone marrow and in the blood circulation.
Discuss the mature erythrocyte, including:
Function and normal lifespan
Membrane characteristics
Energy pathways
Destruction
Objectives
Discuss the spleen’s role in monitoring red cell deformability.
Correlate normal and abnormal RBC size and haemoglobin
content with MCV, MCH, and MCHC values.
Calculate the RBC indices including MCV, MCH and MCHC
Stem Cell Theory of
Haematopoiesis
Pluripotential stem cells give rise to multipotential
progenitor cells
Myeloid stem cell and Lymphoid stem cell
Produce unipotential committed stem cells with receptors
for specific growth factors
Respond by division and maturation (precursor stages)
Erythropoietin-sensitive committed stem cells (CFU-E) red cells
Erythropoietin is lineage specific and regulates RBC production
First recognizable bone marrow precursor is the blast,
following division and maturation of cells
Stem Cell Theory of
Haematopoiesis
Erythropoiesis
Normal RBC and Hgb production requires amino acids,
iron, folic acid, vitamins B12 and B6
Erythropoiesis is controlled by the rate of oxygen delivery
to the tissues
Regulated by erythropoietin (EPO) released by the kidneys in
response to hypoxia; also androgens
EPO stimulates RBC committed stem cells
With normal conditions, 1% of red cells in the blood are
newly released reticulocytes that replace a daily red cell
loss of 1%
Retics remain in the marrow ~2 days before release
Normoblastic Erythrocyte
Maturation
Bone Marrow: Four nucleated RBC stages (1-4)
Marrow & Blood: Reticulocyte (5)
Stage after nucleus extruded; circulates 1 day in blood
9u size, residual RNA called polychromasia on Wright’s stained
smear…amount indicates RBC production rate
Blood: Mature RBC (6)
7u size, fully haemoglobinized, lifespan ~120 days
Six Stages of RBC Maturation Abbott Manual
1 2 3 4 5 6
Normoblastic Erythrocyte
Maturation
Rubriblast Prorubricyte Rubricyte Metarubricyte Reticulocyte Mature RBC
Bone Marrow
NRBC Polychromasia
Mature RBC
RBC Precursors (Stages)
Wright’s stained blood smear
Bone Marrow
Wright’s Stained Blood Smear
Nucleated RBC
Mature Young RBC
RBCs with
Polychromasia
Young RBC
with
Polychromasia
Effective Erythropoiesis
Mature RBC
Functions in oxygen transport and CO2 removal
Lacks a nucleus and organelles
Must maintain membrane integrity
Requires cellular energy and normal haemoglobin
Membrane is a protein shell coated with lipids
Shape essential to oxygen exchange and deformability
Controls volume through control of Na+ and K+
Two major energy pathways
Embden-Meyerhoff ATP, membrane
Hexose-Monophosphate NADPH, keeps GSH reduced
Mature RBC Energy Pathways
Two minor energy pathways
Methaemoglobin reductase Fe+3 Fe+2
Rapoport-Leubering 2,3-DPG
RBC Destruction
Normal red cells circulate ~120 days. With age:
Enzymes are used up
Membrane fragments are lost
Leads to spherocytosis & removal
Normally, ~90% of red cell removal occurs in the
spleen/liver…extravascular haemolysis
Normal deformable RBCs Rigid spherocytes
Spleen Functions
The spleen functions to:
Remove RBC inclusions…pitting
Trap rigid RBCs…culling
Remove excess lipid from retics…grooming
Stores (sequesters)
platelets, ~20%
Filters antigens
Post-Splenectomy Blood
Findings
Target
cell
NucRBC
Howell-Jolly
bodies
Pappenheimer
bodies (target cell)
Howell-Jolly
body
Wright’s stained blood smear
RBC Indices
Define size and/or hgb content of red cells
Used to classify types of anemia and correlate with the
appearance of red cells on blood smear
Indices include:
MCV, mean cell volume, fl
MCH, mean cell haemoglobin, pg
MCHC, mean cell haemoglobin concentration, %
Calculated using RBC, HGB, and HCT values and
reported to nearest tenth
Indices are average/mean values
RBC Indices
MCV = average RBC size, varies with age
Normal adult MCV 82-98 fL = normocytic red cells
MCV <82 fL = microcytic red cells
MCV >98 fL = macrocytic red cells
Normal newborn ~110 fL; normal children ~77 fL
MCH varies with RBC size and Hgb content
MCHC = average hgb concentration per RBC
Normal 32-36% = normochromic red cells
MCHC <32% = red cells may be hypochromic
RDW = index of RBC size variation
Normal 10-14% (low RDW) = uniform RBC size
A high RDW indicates anisocytosis
RBC Indices
Variations in RBC Size and/or
Hgb Content (Color)
Variations in RBC Size and/or
Hgb Content (Color)
Normocytic RBCs Microcytic RBCs Macrocytic RBCs
Small
lymph
Normochromic RBCs Hypochromic RBCs Spherocytes
Evaluation:
3 adults
(1) Normocytic, normochromic anemia, leukopenia, thrombocytopenia
(2) Microcytic, hypochromic anemia, leukocytosis, thrombocytosis
(3) Macrocytic, normochromic anemia (normal WBC & PLT)
Summary
Factors involved in the regulation of erythropoietic activity,
site of synthesis of erythropoietin and its influence on
erythropoiesis.
Stages of RBC maturation, each cell in the RBC
maturation series based on cell size, nuclear size,
cytoplasmic color and nuclear chromatin.
Normal and abnormal RBC size and haemoglobin content
with MCV, MCH, and MCHC values.
RBC indices calculations including MCV, MCH and
MCHC.
