HEMATOLOGY Hematology came from haima and logos Plasma = 55%

LEC 1 BLOOD White blood cells and platelets = <1%

Introduction to Clinical Hematology - nutritive fluid Red blood cells = 45%
Athanasius Kircher (1657) - Participates in the physiologic and pathologic activities of
- 1646 using a microscope: plague victim (Bubonic bacteria) the body SOLID/CELLULAR COMPONENTS
- 1658 Scrutinium Pestis: "little worms" or “ animalcules" Arterial blood – bright red RBC
in the blood Venous blood – dark red - 45% of blood volume
- Proposed hygienic measures: isolation, quarantine, - 4.5 – 6.5 x 1012/L (M/mL)
burning clothes worn by the infected and wearing Functions of Blood WBC
facemasks. - Respiration/transport - Nutrition - 4 – 11 x 109/L (T/mL) - Granular & Agranular
- Excretion Platelets
Anton van Leeuwenhoek (1674) - Homeostasis& -150 – 450 x 109/L (T/mL)
- Father of microbiology - Body protection
- discover microscope GRANULOCYTES & AGRANULOCYTES
Characteristics of Blood Granulocytes
Giulio Bizzozero (late 1800s) - Fluid in vivo - Basophils [0.5 – 1%] → Mast cells - Eosinophils [2 – 4%]
Discovered: - Red in color - Neutrophils [60 – 70%]
- H. pylori - Thick and viscous Agranulocytes
- Function of platelets - Slightly alkaline [pH 7.35 – 7.45] - Lymphocytes [20 – 25%]
James Homer Wright (1902) - Specific gravity = 1.045 – 1.065 - T cells → Th, Tc, Ts, Tm, Tdh
- Pathology Laboratory at the Massachusetts General - 7 – 8% of the total body weight - B cells → Plasma & Memory cell
Hospital - Total volume: [Male = 5-6 L; Female = 4-5 L] - Monocytes [3 – 8%] → Macrophages
- 1902 modification of Romanowsky stain = Wright’s stain
- Megakaryocyte origin of platelets COMPOSITION of BLOOD
LIQUID (PLASMA)SERUM)
Paul Ehrlich ›WATER [91.5%]
- German Bacteriologist ›CHEMICALS
- Classified Leukocytes ›PROTEINS [7%]
›OTHERS [1.5%]
William Hewson ›ELECTROLYTES
- British Anatomist and physiologist - Described Blood ›NPN
coagulation ›HORMONES & ENZYMES
- Isolated fibrinogen ›FOOD MATERIALS
10 Services offered by Hema and Hemostasis Lab • morph – shape • centesis – surgical puncture to remove fluid
- Specimen collection & preparation for exam • myel/myelo – from the BM, spinal cord • algia – pain
- Quantitative manual & instrumental • pan – all, overall, all-inclusive
- Measurements of cells • phleb – vein
- Measurements of cell volumes • phago – eat, ingest
- Evaluation of cellular contents & components • poikilo – varied, irregular
- Cellular identification • schis – split
- Identification of reactive or neoplastic alterations of cell • scler – hard
populations • spleen – spleen
- Evaluation of leukocytes, erythrocytes and platelet • thromb – clot, thrombus
function - Evaluation of cellular development and formation • xanth – yellow
(BM) • peri – around
- Evaluation of hemostatic function • cata – down
• epi – on, over, upon
PREFIXES • anti – antiseptic
• a/an- Lack, without Absent, decreased SUFFIXES
• aniso - Unequal Dissimilar • blast – primitive
• ante – Before • cyte – cell
• Brady – Slow • ectomy – excision, cut out
• Cyto – cell • emia – blood
• Dia – Through • itis – inflammation
• dys – Abnormal/Difficult • lysis – destruction, dissolving
• erythro – Red • logy – study of
• ferr – iron • oma – swelling, tumor
• hemo – pertaining blood • opathy – disease
• hyper – above, beyond, extreme • osis – state, condition, increase
• hypo – beneath, under, deficient, decreased • penia – decrease, lack of
• iso – equal, alike, same • philic – attracted tom affinity for
• leuko – white • plasia – cell production or repair
• macro – large, long • poiesis – cell production, formation
• mal – bad, abnormal • poietin – stimulates production
• mega – large, giant • statis – same, standing still
• meta – after, next, change • trophy – nourishments
• mono – one • spasm – twitch
LEC 2
Safety in the Clinical Hematology Laboratory
Occupational Safety and Health Administration (OSHA)
- US Department of Labor
- Occupational safety and health act (also known as OSHA)
- safe wor
1996 Standard Precautions
- is now used-encompasses Universal Precaution (UP)
and Body Substance Isolation (BSI)
- Unfixed slide – potentially infectious
Potentially infectious materials
- blood
- semen
- all body fluids identified or unidentified
- microhematocrit clay
- unfixed slides
- Direct transmission (injections, syringe) - Indirect
transmission (phone, table tops)
OCCUPATIONAL HAZARDS
1. Biologic Hazards
- exposure to blood and body fluids (greatest risk)
- “Occupational Exposure to Bloodborne Pathogens”
standard (1991)
- Standard precautions protecting lab workers and other
healthcare professionals (March 6, 1992)
Applicable Safety Practices Required by the OSHA
standard
a. Handwashing
- most important safety practice
- Usage of alcogel containing 62% alcohol National Fire Protection Association (NFPA)
- Before and after having a test and getting sample from
patient
- There is visible contamination, removal of gloves
b. Personal Protective Clothing and Equipment (PPE)
- gown must be long sleeved and buttoned
- to avoid spillage
c. Decontamination of work surfaces, equipment
(phlebotomy trays) and spills
- clean after using, to avoid blood contamination
d. Eating, drinking, smoking, applying make-up must
be prohibited in the lab
e. Pipetting method
- no mouth pipetting
- important skill in chemistry
f. Sharps safety and needlestick prevention
- one hand only upon closing the syringe Notes:
g. Hands, pens, fomites must be kept away from - Pull nearest fire alarm
workers mouth and mucous membrane - Call the fire department
- Do crawl to the nearest exit if there is heavy smoke
2. Fire Hazard - Be familiar with Fire Evacuation route, fire blanket, and
- improper use or storage of cryogenic subs (thermal burn) extinguisher location
or subs cable of combustion (fire explosion, or - Don’t panic or run nor use of elevator
asphyxiation)
3. Chemical Hazards
Occupational Exposure to Hazardous Chemical in
Laboratories
Federal register (July 1, 1998)
MSDS – Material Safety Data Sheets (Federal Law)
Ventilations
Labelling – toxic, flammable, carcinogenic
Proper storage – (AAA) Always add acid to water
NFPA Diamond Hazard Symbol LEC 3 Calibrator
Red – Flammability • preserved human/ surrogate cell suspension
Yellow – Radioactivity (Stability standard) Quality Assurance and Quality Control in Hematology • Determined hematologic parameters
White – Special Hazards (Ex. Radioactivity, use no water, 3 Levels of Control
acid) Quality assurance • Normal
Blue – Health Hazards • Coordinated effort to organize lab activities • Normal High
• Provide best service to patients and doctors • Normal Low
Degree of Hazard • Involves control and monitoring Accuracy
4 = extreme hazard ➢ Self-competence • closeness to the true or actual value
3 = serious hazard ➢ Materials and methods Precision
2 = moderate hazard ➢ Reporting of results = TAT ➢ Satisfaction of clients • closeness of results obtained from repeated analysis of
1 = slight hazard ➢ Financial costs same sample
0 = no or minimal hazard Quality Control • reproducibility
1. Procedure Delta Checks
4. Electrical Hazards 2. Evaluate and monitor • assesses change
- Electrical shocks, burns, fire or explosion - Maintenance 3. Specimen of testing system • Compare result from sample analysis with result of
is important 4. Accuracy and precision previous sample
- Be aware of frayed cords, unsafe practices like wet hands Internal Quality Control *same analyte, same patient, dengue patients for in-
on electrical sockets • The actual running of control by the staff stance
• Internal to the laboratory
5. Mechanical Hazards External Quality Control Linearity – the ability to generate results proportional to
- Improper use, storage or disposal of glass wares NEQUAS – National External Quality Assessment Scheme the calculated concentration or activity of the analyte
- shar instruments, compressed gases, or equipment Reliability – extent to which methods is able to maintain
Microhematocrit centrifuge Control accuracy and precision over time
- centrifuges • predetermined Reference Interval
- balanced • same matrix as patient sample • reference range
- lid must never be opened till rotor has completely stopped Primary Standard • range of values of analyte in healthy individuals
• Calibrate instrument Diagnostic Sensitivity- proportion of patients with the
• certified reference material disease with a positive result
• essentially pure from Diagnostic Specificity – proportion of patients identified
• fixed and know composition correctly by the test as not having the disease
Secondary Standard
- analyte concentration ascertained by reference to
10 standard

Systematic Errors - errors within test system or method
Random Errors – occur within prediction or regularity
Preanalytical
• Specimen obtained from wrong patient
• Specimen procured ate the wrong time
• Specimen collected in the wrong tube
• Blood specimen collected in wrong order • Incorrect
labelling
• Improper processing of specimen
Analytical
• Oversight of flagging
• Out of control quality control results • Wrong assay
performed
Postanalytical
• Verbal reporting of results
• Lab Information System incompatibility error
• Confusion about reference ranges
• Failure to report critical values ASAP
LEC 4 SPECIMEN TRANSPORT- proper handling and timely
SPECIMEN COLLECTION VENIPUNCTURE transport
- Sources of Error (refer to page 17 Koepke table 2-5)
PHLEBOTOMY VENIPUNCTURE Common during venipuncture procedure are
- Most common technique used to obtain blood. - failure to dry site
- Requires ample skill to ensure accurate results and - bevel down,
preservation of patient vein integrity - failure to mix blood ASAP
Initial steps - failure to release tourniquet
Correct Patient Identification - excessive pulling of plunger.
1st critical step (compare info in the lab request) (verify the
age and sex) SPECIAL SITUATIONS encountered
- Note Isolation Restrictions IV site/Med lock- never drawn on the same side
- Note dietary restrictions - opposite arm
- Reassure the patient-don’t deceive - distal or below IV line (noted)
- Position the patient-lie in bed for (admitted patient) for Transfusion
ambulatory patient - extraction chair - may collect blood.
- Assemble supplies-accessibility - should be indicated and drawn opposite arm.
- Selection of the site: median cubital (preferred) median HD shunt/Fistula
basilic, cephalic veins. If not veins of the wrist, back of - avoided.
the hand, ankle or foot (consult if there is circulatory Ask for the status may draw distal at least 4 inches below.
problems). Mastectomy
Points: avoid areas w/ hematoma, burns, scars or edema, - collected on the other side. Mastectomy promotes
IV-line, mastectomy site also. lymphostasis
- Apply the tourniquet-3-4 inches above the site. - Clean Hematoma, Burned or Scarred areas- avoided.
the site properly - Scarring causes veins difficult to palpate
- Inspect the needle and the vacuum Burned areas are prone to infection
- Release the tourniquet-always before removal of needle
- Remove needle and apply pressure- do not allow patient REMEMBER: each phlebotomist is allowed only of two
to bend the arm this will reopen the wound and result to attempts.
hemorrhage.
- Discard needle
LABEL SPECIMEN- no pre-labelling
(name, date and time, age/sex, initials of phlebotomist and
any info required by the institution)
 SPECIMEN COLLECTION SKIN PUNCTURE TECHNIQUES IN PERFORMING CBC
- Done in infants particularly newborns
- (hospital induced anemia), osteomyelitis-puncture not
more than 2.4mm
- Safer
- Puncture site should be warm
Sites of skin puncture:
- 3rd or 4th finger
- big toes
Manual – 2 to 3 times = 100 WBC to identify
- heel
Machine – release result less than 1 minute in one patient
- last resort is the earlobe
Reasons for skin puncture
Hemoglobin (Hb)
- For adults because of obesity, burns, extremely small
Estimated by:
veins
LEC 5 - color
- Blood is a mixture of capillary, venous and arterial blood
BASIC HEMATOLOGICAL TECHNIQUES - power of combining with oxygen or carbon dioxide - iron
w/ interstitial and intracellular fluid
content (iron deficiency)
- It is important to wipe the first drop of blood
Complete Blood Count (CBC) Detection and assessing clinical anemia
- A good puncture requires no forcing or hard squeezing.
- Foundation procedure Hemiglobincyanide (HiCN) or Cyanmethemoglobin
- Most common method (Manual) (Dragkin’s reagent)
Sources of error
- Hemolysis - Disorders (Hematologic or hematologic manifestations
secondary to other diseases) abnormalities in RBCs, Note: 1 gram of hemoglobin can carry 1.34 ml of oxygen
- Failure to dry the site
WBCs and/or platelets Acidhematin method - alternative laboratory method
- Failure to wipe the 1st drop of blood
Ex. Anemia (manifestation) Reagent: 0.1 normal hydrochloric acid
- Vigorous massaging or milking the area - Accidental
capturing of bubbles
Components REFERENCE VALUE
1. Hemoglobin (Hb) Reference values of hemoglobin Common unit = g/dl
2. Hematocrit SI unit = g/L (x10)
3. RBC count with morphology (Manual RBC is not
performed now a days)
4. WBC count with differential (manual or automated)
5. Platelet estimate (manual method)
Platelet count (machine) 150,000-450,000
6. RBC indices
Hematocrit  MCV Platelets
 Packed cell volume - Volume or size of average RBC Thrombocytosis (high) or Thrombocytopenia (low)-
 Simple screening test for anemia Reference values of MCV: 80 - 100 fL (femtoliter) Associated with hemostatic mechanism
 Reference method for calibrating blood count systems Formula of MCV: Reference values:
 Rough guide to accuracy of Hb measurements 150 – 450 x 10^9/L Leukocyte Differential Count
 Used in the calculation of red cell indices MCV= Hematocrit x 10/RBC count = Femtoliter
 Absolute (x 10^9/L) = multiply relative number of WBCs
Reference values of hematocrit e.g. 450 x 10/5.12 = 87.89 (normal) by the total WBC count/L
Common unit = Percent  MCH
SI unit = L/L - Weight of hemoglobin in average RBC Reticulocyte Differential Count
Reference values of MCH: 27 – 33 pg (picogram) - Juvenile red cells demonstrated by Supravital stains -
Formula of MCH: Reflects erythropoietic activity of Bone marrow
MCH = Hemoglobin x 10/RBC count = picogram
Ex. 14.2 x 10/5.12= 27.73 (normal) Relative count (percentage)
PS: gi minus ug plus lang na siya, - Adult: 0.5 – 1.5%
0.47- 0.07= 0.40 0.47+ 0.07 = 0.54  MCHC - Neonates: 1.5 – 6.5%
- Hemoglobin concentration or color of the average RBC
Rule of three Reference values of MCHC: 32 – 36 g/dL Absolute count (x10^9/L)
- 3 x RBC = Hemoglobin + 0.5 Formula of MCHC: = 5.0 x 10^9/L
- 3 x Hemoglobin = Hematocrit + 3% Only apply to red MCHC = Hemoglobin x 100/Hematocrit= g/d
cells e.g. 12 x 100/45 = 26.66 g/dL (below)
hypochromic- pale
Red Cell Indices Blood Cell count hyperchromic- high hemoglobin content
- Morphologic classification of anemias normochromic- normal
- Used also in QC RBC microcytic- small
- Calculated from Hb, Hct, and RBC count Anemia (High) or Polycythemia (Low) macrocytic- big
Reference Values: normocytic- normal
3 indices Male: 4.6 – 6.0 x 10^12/L
- MCV (Mean Corpuscular Volume) Female: 4.0 – 5.4 x 10^12/L
- MCH (Mean Corpuscular Hemoglobin)
-MCHC (Mean Corpuscular Hemoglobin WBC
Content/Concentration) Leukocytosis (high) or Leukopenia (low) - Indicate infection,
follow disease progress
Usually used is MCV and MCHC Reference values: 4.5 – 11.5 x 10^9/L (rodak)
ABSOLUTE COUNT  HEME VARIATIONS in VALUES
-iron-containing
Total WBC = 5.0 x 10^9/L  GLOBIN • Altitude
Differentials: -Heme group • Age
Neutrophil – 75% - 0.75 x 5.0 = 3.75 -protein-portion - birth (18-21.5 g/dL)
Lymphocyte – 20% - 0.20 x 5.0 = 1.0  OXYHEMOGLOBIN - infancy (10-14 g/dL)
Eosinophil – 4% - 0.04 x 5.0 = 0.2 - saturated with oxygen - 10 years to adult (12-15 g/dL)
Monocyte – 1% - 0.01 x 5.0 = 0.05  DEOXYHEMOGLOBIN • Gender
100% 5.0 - no bound oxygen in complex with carbon dioxide - Male (14-18 g/dL)
- Female (12-15 g/dL)
Erythrocyte Sedimentation Rate (ESR) HEMOGLOBIN VARIANTS
- Measure of degree of settling of RBC in plasma in an GRAVIMETRIC METHOD
anticoagulated whole blood sample during specified period NORMAL • Not routinely done
of time.  HbA • Blood donor screening
 HbA2 • A drop of blood is allowed to fall into a copper sulfate
Two Methods:  Hb F (1.053)
1. Westergren • RESULTS:
2. Wintrobe method ABNORMAL - Sink = >12.5 g/dL
 HbS - Float =<12.5 g/dL
Normal values: Westergren  Hb C
0–50 years old (male) = 0.15 mm/hr more than 50: 0-20 0-  Hb D CYANMETHEMOGLOBIN METHOD
50 years old(female)= 0.20 mm/hr more than 50: 0-30  Hb E • most widely used method
• Reagent: DRABKIN'S REAGENT
HEMOGLOBIN DERIVATIVES Hgb + Drabkin's rgt = CYANMETHEMOGLOBIN
 OXYHEMOGLOBIN
LEC 6  DEOXYHEMOGLOBIN
 CARBOXYHEMOGLOBIN
HEMOGLOBIN DETERMINATION  METHEMOGLOBIN
 CYANMETHEMOGLOBIN
HEMOGLOBIN  SULFHEMOGLOBIN
• a tetramer composed of two identical globin chains, each
of which binds a heme molecule
•Primary constituent of red blood cell cytoplasm and
transports molecular oxygen from the lungs to the tissues
ACID HEMATIN METHOD 2. Pluripotential- present several days after fertilization.
• Reagent: 0.1 N HYDROCHLORIC ACID Can develop into any cell type except into being a
• Sahli-Hellige hemoglobinometer set fetus.
Hemoglobin + 0.1 N HCI = ACID HEMATIN 3. Multipotential- derived from pluripotent cells. Found
in adults and are limited to specific types of cells to
AUTOMATED METHOD form tissues.
• Convenient and safe HEMATOPOIETIC DEVELOPMENTAL PERIODS
• modified HiCN method
• cyanide reagent or sodium lauryl sulphate 1. Mesoblastic Period (yolk sac hematopoiesis)
• altered concentrations of reagents, temperature, and  19-20 days of gestation (2nd week)
pH of reactions  Occurs in blood island of the yolk sac
• non-ionic detergent  Primitive erythroblasts
 Found in yolk sac arising from mesodermal
LEC 7 cells.
 Occurs intravascularly
HEMATOPOIESIS  Angioblasts- forms future blood vessels
 the process or continuous as well as regulated PERIOD
process of blood cell production, differentiation and Portland= 2 zeta & 2 gamma chains
development and maturation Gower I= 2 zeta & 2 epsilon
 Restricted in bone marrow Gower II= 2 alpha and 2 epsilon
ORIGIN OF BLOOD CELLS 2. HEPATIC PERIOD
 Hematopoietic Stem Cells (HSCs) foundation of  4th to 5th or 5th to 6th week of gestation
adult hematopoietic system  definitive morphologic hematopoiesis
 Now widely accepted that it was produced by -presence of erythroblasts + lymphoid cells
embryo  Fetal liver:
 Repopulation -primary erythroid organ (extravascular)
Spleen- involved solely in lymphopoiesis
TYPES OF HUMAN STEM CELLS
Starting of megakaryocyte production
1. Totipotential- present in the first few hours after an Fetal hemoglobin (common Hb) – 2 alpha 2 gamma
ovum is fertilized. The most versatile. Hb A1 and Hb A2 also detected.
 Organs involved: Spleen, thymus, and lymph
nodes
 Fetal hepatic hematopoiesis => Adult extramedullary
hematopoiesis => Bone marrow failure (rare) (liver
reduce blood cells)
3. Myeloid/Medullary Period
 5th month of gestation
 Developing bone marrow cavity (long-bones) inner
part of Bone marrow
 In end of 6th month, bone marrow is the main site
for hematopoiesis replacing the fetal liver.
 Production of EPO (erythropoietin), G-CSF,
GranMacro-CSF, HbF, Hb A2 and Hb A1
 4th year of life = marrow replaced by fats
(hematopoietically inactive marrow)
ADULT HEMATOPOIETIC TISSUE
A. BONE MARROW
Two types of Bone Marrow
1. Red Marrow (Sternum, Scapula, Clavicle, Femur,
Skull, Ribs, Spinal column and Pelvic bone)
 Hematopoietically active
 Flat bones and proximal ends of long bones
2. Yellow Marrow
 Hematopoietically inactive
 Adipocytes (fat cells)
 Normally – 50% red marrow and 50% yellow
marrow
 Retrogression
 process of replacing red marrow by yellow marrow Culling Pathophysiology:
starting 4th year until 7th year of life  Cells are phagocytose with subsequent
degradation of cell Adenitis
B. LIVER Pitting  Infection of lymph nodes
 Major site in hepatic hematopoiesis (fetus)  Removed damage from circulating red cells Metastasis
Hematopoietic functions (adults):  IgM synthesis  Infiltration by malignant cells
 Synthesis of various transport CHON  Storage for platelets (1/3) stored in spleen
 Storage of Minerals and Vitamins
 Bilirubin Conjugation E. THYMUS
 Bilirubin transportation  Efficient, well developed at birth
Pathophysiology:
Pathophysiology: Pathophysiology:
Hemolysis
Porphyrias  Increased environmental stress due to small Non-dev’t (gestation)
 Enzymatic deficiencies openings by inter endothelial junction  Lack of T-lymphocytes
 Accumulation of intermediary porphyrins Splenomegaly  Uncontrollable infection death
 Enlarged, palpable Thymic disturbance (adult)
Bilirubin conjugation Fe ++ - storage Hypersplenism  no effect
 Severe H.A. & RBC dysplasia  Enlarged leading to Pancytopenia
 Caused commonly by congestive splenomegaly STEM CELL THEORIES
Storage diseases secondary to liver cirrhosis and portal
 Monocytes/macrophages (Kupffer cells) hypertension PLURIPOTENTIAL STEM CELL
 Enzymatic deficiencies  One cell gives rise the varied blood cells (E, L, M)
 Hepatomegaly with liver dysfunction D. LYMPH NODES  Widely accepted theory
 Bean shaped Member of lymphatic system
C. SPLEEN occurring in groups or in chains and are located COMMITTED STEM CELLS
 Largest lymphoid organ superficially or deep.  Give rise to descendants or progeny cells that
 Vital but not essential for life eventually become restricted to a specific cell line
Functions: development
Functions:  Formation of new lymphocytes
 Indiscriminate filter of blood  Processing of specific Immunoglobins Characteristics/ Fates of Stem cells
 Sequesters senescent red cells  Filter particulate matter, debris, bacteria entering  Capable of self-renewal
 (Old, 350 ml 120 days) via lymph  Give rise to differentiated progeny
  Able to reconstitute the hematopoietic system of  Stimulating hormonal factor is PO which is mainly
an irradiated host Leukopoiesis produced by the liver.
 Apoptosis  Colony Stimulating Factors
 Interleukins (IL#)
CYTOKINES (hematopoietic growth factors)
 Group of specific glycoCHON Megakaryocytopoiesis Lec 8
 Regulates the proliferation, differentiation, and  Thrombopoietin (hormone)
maturation of hematopoietic precursor cells  Meg-CSF NORMOBLASTIC MATURATION
 Include interleukins, lymphokines, monokines, Erythropoiesis
chemokines, interferon, and colony stimulating  Occurs in Bone marrow  Red Blood Cells
factors  CFU-GEMM gives rise to earliest identifiable colony of  called erythroctes
RBC- Burst Forming Unit- Erythroid (BFU-E)  Its 1 true fxn-carry oxygen from the lung to the
Colony Stimulating Factors (CSFs)  BFU-E will become CFU-E that has many EPO tissues
 Produced by many cells receptors - EPO induces Hb synthesis  There are 3 nomenclatures in naming RBC
 High specificity to target cells precursors.
 Active at low concentrations Leukopoiesis 1. Erythroblast terminology- Europe
 (eg: G-CSF)- primary target is granulocyte Major Categories 2. Normoblastic terminology- USA
 Can also have synergistic effect (IL-3 and G-CSF for 1. Macrophage 3. Rubriblast terminology- used by some since it
Megakaryocyte colony stimulation) 2. Lymphopoeisis parallels the WBC development nomenclature

Interleukins GM-CSF, G-CSF, M-CSF including IL-3, IL-5, IL 11 – B Nomenclature for Erythroid Precursors
 Numbered in order of identification cell
 CXS:  B cell growth factor Normoblastic
 CHON that exhibit multiple biologic activities IL-3-multilineage: stimulating granulo, mono, mega,  Pronormoblast
synergistic interactions with other cytokines and and erythroid cells  Basophilic Normoblast
growth factors  T-cell  Polychromatic (Polychromatophilic) Normoblast
 Interacting systems with amplification potential  IL-2  Orthochromic Normoblast
 Effective at very low concentrations  Reticulocyte
Megakaryopoiesis  Erythrocyte
Factors that stimulate linage (specific hematopoiesis)  Platelets
 Involved in hemostasis and thrombus devt. Rubriblastic
Erythropoiesis  Earlier influences include GM-CSF, IL-3, IL-6, IL-11, kit  Rubriblast
 Erythropoietin (hormone= produced by kidneys) ligand and EPO  Prorubricyte
 Hypoxia (low oxygen)  Rubricyte
  Metarubricyte  Size: 8 um
 Reticulocyte  Basophilic Normoblast  Cellular activity: continuation of, Howell-Jolly
 Erythrocyte  Nucleus: Ratio (6:1), chromatin begins to Body production
condense, staining results is deep purple red  Length of time in this stage: approximately 48
Erythroblastic  Cytoplasm: deep blue hrs
 Proerythroblast  Division: mitosis
 Basophilic Erythroblast  Location: only in BM  Polychromatophilic Erythrocyte/Reticulocyte
 Polychromic Erythroblast  Size: 16um  Nucleus: none
 Orthochromic Erythroblast  Cellular activity: detectable HB synthesis  Cytoplasm: predominantly the color of HB
 Reticulocyte occurs  Division: none
 Erythrocyte  Length of time in this stage: slightly more than  Location: in BM for 1-2 days then peripheral
24 hrs. blood 1 day
Maturation Process Erythroid Progenitors  Size: 8 um
 BFU-E (burst forming unit-erythroid) 1 week to mature  Polychromatic/polychromatophilic Normoblast  Cellular activity: completes the HB production
 CFU-E(colony forming unit- erythroid) 1 week also  Nucleus: ratio (4:1), chromatin condensation, and endoribonuclease digests the ribosomes
towards becoming the pronormoblast no nucleoli  Length of time in this stage: 2-8 days
 Approximately 6 days for precursors to mature  Cytoplasm: 1st stage Redness/pink is
 It will take approximately of 126 days to produce a associwith HB, and concurrent decrease of  Erythrocytes
RBC RNA. Mixed pink and blue (murky gray-blue)  Nucleus: none
 Division: mitosis( last stage capable)  Cytoplasm: mature cells are biconcave discs.
Maturation Sequence  Location: in BM Salmon-pink in color when stained with a
 Pronormoblast,  Size: 13um central area (concavity)
 Nucleus: high (N8:C1), round to oval with 1 or 2  Cellular activity: HB synthesis is increasing &  Division: can't divide
nucleoli the accumulation is visible in the cytoplasm.  Location & Length: active approx. 120 days
 Cytoplasm: quite blue (ER), Golgi complex may Organelles still present  in the circulation
be visible  Length of time in this stage: 30 hrs.  Size: 6-8 um
 Division: Mitosis  Shape: biconcave
 Location: in BM  Orthochromic Normoblast  Cellular activity: delivers oxygen to tissues,
 Size: 18-20 um  Nucleus: completely condensed or nearly so. releases it, and returns to the lung to be re-
 Cellular activity: accumulate components for HB Low Nucleus to cytoplasm ratio 1:2 oxygenated.
production and globin production begins  Cytoplasm: pink-orange/salmon pink color
 Length of time in this stage: slightly more than 24 (nearly complete HB production)
hrs.  Division: none
 Location: in BM