GENERAL CHARACTERISTICS:

 A liquid form of CT in which cells are suspended in a circulating fluid or plasma (intercellular substance).  Structurally & genetically related to CT since it also arises from mesodermal mesenchyme.  In man, its volume is about 5l, constituting about 7% of the body weight.

 When anticoagulants are added, blood samples can be centrifuged into 3 major fractions: 1.Sedimented RBC- constitutes 43% of total blood volume (hematocrit is used to refer to the estimate of the volume of packed RBC per unit of blood). 2.Buffy coat - a thin layer of WBC between the RBC layer and supernatant light-colored plasma layer 3.On the upper surface of the buffy coat is a thin layer of platelets.

 Knowledge of normal histology of blood is important for diagnostic purposes: e.g. anemia, leukemia, viral and parasitic infections.  Blood cells and their components exhibit 4 major staining responses to Romanovsky-type mixtures (Giemsa or Wrights stain):  Basophilia- affinity for methylene blue; stain purple to black  Azurophilia- affinity for oxidation products of methylene blue called azures; stain red-blue  Eosinophilia- affinity for eosin; stain orangepink to orange  Neutrophilia- affinity for a complex of dyes; stain salmon pink to lilac

1.PLASMA- transparent yellow but becomes milky opalescent after a meal due to minute droplets of absorbed lipid in it. The contraction of clotted blood or plasma expresses a clear, yellowish fluid the serum, which has no more fibrinogen content. 2.SOLID OR FORMED ELEMENTS- RBC, WBC, blood platelets 3.NON-CELLULAR ELEMENTS: Chylomicrons (tiny fat droplets carried to the tissues as nutrition); Hemoconia or blood dust (cellular debris of destroyed RBC) 4.LYMPH- a colorless fluid which is collected from the tissues and returned to the bloodstream.  Lymphocytes and IgGs are added as lymph passes through lymphoid organs.  Rich in fat globules and glycogen; milky lymph from the small intestine is referred to as chyle.  Coagulates with a soft clot but more slowly than in the blood.  Circulates in only one direction- towards the heart.

 Most numerous among the formed elements (5.4 million/cc in men; 4.8 million/cc in women).  Are biconcave, non-motile disks of around 7.5 microns in diameter, elastic & are capable of considerable distortion.  The life span as determined by differential agglutination is generally from 30-120 days, after which the cells are destroyed by reticulo-endothelial cells of the spleen & liver.  Are not true cells as they lack a nucleus, mitochondria and other organelles. They develop in the bone marrow but during the final stages of maturation, the nucleus and other organelles are extruded.  Specialized for oxygen transport from lungs to tissues, and return of CO2 to the lungs.

 The major integral membrane glycoprotein of RBCs is glycophorin.  Enzymatic systems & antigens for ABO major blood groups & Rh factor are built into this membrane.  The membrane skeleton is composed of spectrin, actin and associated proteins (tropomyosin, ankyrin, band 3, band 4.1, band 4.2, 4.9).  Spectin deficiencies or defective binding of spectrin to integral proteins result to hereditary diseases known as spherocytosis and elliptocystosis.

 A fresh erythrocyte is pale, greenish yellow in color, homogenous, and nongranular. In dense masses, the color turns red.  In fresh blood smears or stagnant circulation, RBC form long columns termed Rouleaux formation. This maybe due to surface tension.  In pathological conditions, RBC may agglutinate (sludging or clumping) due to agglutinins forming the blood groups.

 The interior of the cell contains a 33% solution of the O2 and CO2 carrying protein hemoglobin that accounts for their acidophilia. In addition, there are enzymes of the glycolytic and hexose-monophosphate-shunt pathways of glucose metabolism.  When hemoglobin content is increased (normochromic12-15 g/cc of human blood) the cells become darker (hyperchromic). When scanty, the cells stain paler with a bigger halo in the central portion (hypochromic).  Combined with O2 or CO2, hemoglobin respectively forms oxyhemoglobin or carbaminohemoglobin.  Carboxyhemoglobin causes a reduced capacity of blood to transport O2.

 Normal adult hemoglobin (HbA: 2E, 2F chains) constitutes 96% of hemoglobin; fetal hemoglobin (HbF: 2E, 2K chains) is less than 2%. The latter is abnormally persistent in a kind of anemia called thalassemia.  HbS is the abnormal form of HbA in sickle-cell anemia which is a fatal point mutation disorder where RBC becomes inflexible and fragile, easily bursts, clogs and retards blood flow in capillaries, leading to severe anoxia in tissues.  Residual hemoglobin is processed by the liver and excreted as the pigment bilirubin.

 In stained specimens (Giemsa stain), the cytoplasm appears reddish or orange- pink in color with a small pale-staining central area due to lesser hemoglobin.  No organelles are present. The cytoplasm is nongranular, but sometimes show polychromatophilic particles which are small, angular basophilic structures (possibly remnants of the nucleus) referred to as Howell-Jolly bodies.  The contents of the corpuscle are normally in osmotic equilibrium (isotonic) with the plasma. In hypertonic solution the cell (echinocyte) loses water, so that the outlines becomes scalloped with spiny projections (crenation). A hypotonic solution cause swelling followed by destruction and liberation of hemoglobin (hemolysis). The cellular debris gives rise to shadow cells or erythrocyte ghosts, which finally break into tiny particles called blood dust or hemoconia.

 Reticulocytes are immature RBC which appear larger and exhibit a slight bluish stain because of the presence of residual RNA. Reticulocyte count is a rough index of erythrocyte formation; elevated counts in anemia is a sign of response to treatment.  Anisocytosis (variation in size) are observed in different pathological conditions: microcyte (smaller than normal size); macrocyte or megalocyte (bigger). In poikilocytosis (variation in shape), RBC may appear oval or sickle-shaped.  A decreased number of RBC is usually associated with anemia, characterized by lowered blood concentrations of hemoglobin (hypochromic anemia). Anemia maybe caused by loss of blood, insufficient production of RBC by bone marrow, production of RBC with low hemoglobin (due to iron deficiency), or accelerated destruction of blood cells.  Polycythemia (increased number of RBC) occurs during ascent to high altitudes, or in diseases characterized by an increased hematocrit, resulting to increased blood viscosity.

Spherocytosis

Thalassemia major

Pernicious anemia Sickle-cell anemia G6PD deficiency

Lead poisioning

Fe+2-deficiency anemia

Howelll-Jolly bodies

LEUKOCYTES (WHITE BLOOD CORPUSCLES)

 Nucleated, spherical, motile cells with no hemoglobin content and no distinct cell membrane.  The cytoplasm contains two types of granules: specific granules with specific functions bind neutral or acidic components of the dye mixture, and azurophil granules that stain purple and are primary lysosomes.  The normal count ranges from 5,000-9,000/cc of blood. In the presence of acute infections, WBC count may rise to 20,000-40,000/cc (leukocytosis); a decreased count is termed leukopenia.  They are amorphous, and are capable of penetrating between endothelial cells into CT by means of a process called diapedesis.  They remain in circulation for only about 24 hours. Billions of granulocytes die by apoptosis each day in adults; resulting debris is removed by macrophages without eliciting an inflammatory response.

1.GRANULOCYTES or POLYMORPHONUCLEARScontain membrane-bound granules with hydrolytic action; hence belong to the class of lysosomes; nuclei are lobated, segmented, or polymorphous. They are incapable of mitosis. 2.AGRANULOCYTES- instead of specific staining granules, they have very fine azurophilic or metachromatic granules; size varies from 7-17 microns.

GRANULOCYTES
1.NEUTROPHILS- 55-65% of the total WBC count.  Are 10-12 mm in diameter. Normal life span is about 8 days, mainly spent in reserve in the bone marrow.  Nucleus consists of 2-5 lobes (polys) connected by thin strands of chromatin. Hypersegmented cells (more than 5 lobes) are typically old neutrophils.  A drumstick nuclear appendage (Barr body) in females represent the condensed X chromosome.  Contain glycogen for energy production. Failure of oxidative metabolism in neutrophils account for dark color of necrotic tissues.

 The finely granular cytoplasm stains lilac or purple. Specific granules greatly outnumber azurophil granules. The former lack lysosomal enzymes but contain alkaline phosphatase and basic antibacterial proteins (phagocytins).  As phagocytes (microphages), they have a powerful microbial killing system: an acidic environment, and production of hypochlorite, superoxide (O2-), H2O2-, myeloperoxidase and halide ions.  They constitute the 1st line of body defense against invading organisms, but are not equally effective against all types of bacteria. Dead neutrophils constitute pus which accumulate in sites of infection.

In immune phagocytosis, an antibody (IgG) binds to the surface of the bacterium, & a complement component of the plasma (C3b) binds to the antigen-antibody complex. Receptors in the neutrophil membrane promote the adherence of these opsonin complex to its surface, increasing phagocytosis efficiency. Nonspecific phagocytosis do not employ opsonins.

 To combat a bacterial invasion, neutrophils and endothelial cells both release substances at sites of infection: LCAM-1 (leukocyte adhesion molecule), and cytokines such as IL-1F, tumor necrosis factor (TNF-E) that stimulate the endothelium to synthesize the endothelial leukocyte adhesion molecule (ELAM-1).  They also generate leukotrienes (LTs) which serve various functions: promote their adhesion to endothelium, as chemoattractants for other WBC, contribute to edema at sites of inflammation, and as vasoconstrictors.  Drugs blocking the synthesis of leukotrienes are effective in the treatment of asthma.

2.EOSINOPHILS- 1-3% of the WBC count  Nucleus usually has 2 oval lobes connected by a nuclear thread; there is no nucleolus.  Coarse specific granules stain pink with Wrights stain and contains proteins not found in other cell types: the major basic protein, and the eosinophilic cationic protein which have cytoxic effects on parasitic worms; and eosinophil-derived neurotoxin, the function of which is not well understood.

 Like neutrophilic granules, they contain lysosomal enzymes but have a higher concentration of peroxidases; they lack phagocytins.  They are not avid phagocytes, but can phagocytose antibody-antigen complexes.  They produce substances that modulate inflammation by inactivating the leukotrienes and histamine produced by other cells.  Elevated circulating counts are observed in allergic and hypersensitivity conditions like asthma, parasitism and certain skin diseases.

3.BASOPHILS- constitute only about 0.5-1% of the WBC count, being increased around cancer growths, in leukemia and chicken pox.  The heterochromatic nucleus is divided into irregular lobes obscured by overlying basophilic specific granules.  Basophils supplement functions of mast cells in immediate hypersensitivity reactions since they also elaborate heparin and histamine from their granules in response to IGEs.  Basophils and mast cells are not the same, since they are ultrastructurally (mast cells are larger, with round nuclei, abundant granules, sessile and relatively long-lived), and genetically different.

AGRANULOCYTES
4.LYMPHOCYTES- 2nd most abundant class of WBC, constituting 20-35% of circulating blood cells. They are primarily concerned with the immunological responses of the body.  Slightly larger than RBC (7-8 microns in diameter), spherical cells with a big, intense-staining round or slightly indented nucleus surrounded by a thin rim of clear blue cytoplasm.  Has Golgi complex, a pair of centrioles, and very few mitochondria, no ER. Azurophilic granules are seen only in small lymphocytes.

Types of lymphocytes: a. B-lymphocytes (bursadependent) - move directly to lymphoid tissues via the bloodstream. They survive only for a few days and are involved in humoral immunity. b.T-lymphocytes (thymusdependent or memory cells)responsible for cellular immunity.  Re-enter the bloodstream from the thymus and return to the bone marrow or lymphoid organs where they live for months or years. c. Null (NK) cells- morphologically resemble lymphocytes but are neither B or T cells. They may represent circulating stem cells or other blood cell types.

 Possess peripheral E,F or K, H heterodimeric surface antigen receptors.  The former interacts with peptides of class I or class II MHC (major histocompatibility complex) molecules; the specificity of the latter is still unknown.  Functions include: acting upon foreign or abnormal materials, producing a delayed hypersensitivity reaction, and regulating behavior of Blymphocytes through helper and suppressor T-lymphocytes.

 Cytotoxic or killer Tlymphocytes are the agents behind cell-mediated immunity, & are responsible for graft rejection after organ transplantation.  Upon recognition of their target cells, the killer cells synthesize perforin , a pore forming protein that creates pores leading to cell lysis.  To activate T-cells, antigen must be presented together with proteins coded by MHC genes of the host. This process is augmented in the presence of macrophages, and mediated by signaling molecules called lymphokines.

5.MONOCYTES - constitute 3-8% of the leukocytes; more abundant in later stages of TB and malaria.  Resemble the large lymphocytes except that the cytoplasm is more abundant & has a grayish-blue tint with scattered azurophil granules.  There are conspicuous Golgi complex, scanty rER, and some glycogen granules.  The slightly indented and vesicular nucleus is eccentrically located. 2 or more nucleoli are seen.  They constitute a mobile reserve of scavengers and are essential for the processing of antigens prior to the development of antibodies by B-lymphocytes.  CT macrophages (histiocytes) and monocytes are identical cell types.

6.BLOOD PLATELETS are small, colorless, nonmotile, non-nucleated and slightly basophilic cellular elements; varies from 200,000-400,000/cc of blood.  Lightest (2-4 microns) elements of blood, adhering easily to the surface they come in contact; life span is approximately 10 days.  The blood of reptiles, birds and amphibians have nucleated cells called thrombocytes that have the same role in blood clotting as the platelets.  Are cytoplasmic fragments of giant cells or megakaryocytes in the bone marrow.

Each platelet consist of 2 concentric zones under the LM: a. Chromomere or granulomere- the thicker central area contains small purple-staining granules resembling a nucleus. 2 types are present: alpha granules contain fibrinogen, platelet-derived growth factor, other proteins; delta granules contain serotonin; lambda granules contain lysosomal enzymes. b.Hyalomere- thin, non-granular homogenous peripheral zone surrounding the chromomere.  It is free of granules but has fine filaments of microtubular bundles that help maintain the discoid shape of platelets. An actomyosin complex (thrombostenin) constitute 15% of total platelet protein, allowing them to become contractile during platelet movement and aggregation.

Their principal functions include:  Primary aggregation- patch small defects in the endothelial lining of blood vessels by aggregating on damaged tissue, forming a platelet plug.  Secondary aggregation- platelets release the content of their alpha and delta granules. Serotonin, ATP, ADP are all potent inducers of platelet aggregation.  Blood coagulation- factors from the blood plasma, damaged blood vessels, and platelets promote cascade interactions of approximately 13 plasma proteins.  The cascade can be initiated by 2 converging pathways, each of which results in the formation of a thrombus (blood clot) through the conversion of fibrinogen to fibrin by the enzyme thrombin.

1.In the intrinsic pathway, factor XII is activated by contact with collagen underlying the damaged endothelium 2.In the extrinsic pathway, tissue thromboplastin (factor III) combines with calcium and factor XII to activate factor X, a plasma protein. Factor X is the point of convergence of the 2 pathways, and in its activated form, promotes the conversion of prothrombin (factor II) into thrombin, which in turn transforms fibrinogen to the clot polymer fibrin.

 Both factor X and prothrombin are synthesized by the liver and require vitamin K as a cofactor in their synthesis.  Other factors such as platelet cyclo-oxygenase enzymes convert prostaglandin endoperoxides to thromboxane that in turn accelerate blood clotting.  Aspirin is known to inactivate these enzymes and is clinically used to diminish the risk of coronary thrombosis.  Clot retraction and removal- interaction of platelet contractile proteins thrombosthenin, actin, myosin and ATP contracts the clot, which is then removed by lambda granules, and the proteolytic enzyme plasmin, formed through the activation of the plasma proenzyme plasminogen, by endothelium-produced plasminogen activators.

 Human platelets contain platelet-derived growth factor (also synthesized by monocytes, macrophages and endothelial cells, stimulate fibroblast proliferation for repair of damaged blood vessels), von Willebrands factor (promote adhesion of platelets to endothelial cells), platelet factor IV (stimulates blood coagulation), and thrombospondin (involved in platelet aggregation).  Inherited abnormalities of platelet or deficiency of factors include:  hemophilia (bleeding disorder due to absence of blood clotting factors)  thrombocytopathia (abnormalities in platelet structure and function)  thrombocytopenic purpura (characterized by platelet deficiency, and abnormal fragility of small blood vessels that lead to black and blue areas over body surface).

inability to form a blood clot

HEMOPOIESIS or HISTOGENESIS OF BLOOD CELLS

 Mesoblastic phase- blood formation in the body stalk and yolk sac. Mesenchymal cells form blood islands and differentiate into primitive erythroblasts and nucleated erythrocytes.  Hepatic phase- cells called definitive erythroblasts appear in the liver primordium; granulocytes and megakaryocytes also appear in the sinusoids in the 2nd month. Later, the spleen also become a hemopoietic site.  Myeloid phase- begins with the establishment of ossification centers in cartilage of long bones during the 4th-5th month of intrauterine life. As the marrow becomes established in the bones, hemopoiesis in the liver and spleen declines.

 Blood cell formation occurs both in embryonic life and also in certain pathological conditions:  Homoplastic or constitutive hemopoiesis occurs when blood arises by division of pre-existing immature cells of the series.  Heteroplastic or induced hemopoiesis occurs when there is an increased demand for blood, so cells are formed directly from stem cells called hematocytoblasts.
Lymphoid tissue of liver

Bone marrow a highly specialized adult reticular CT with hematogenic and osteogenic potential.

1.Red or non-fatty bone marrow (myeloid tissue) is found in bone cavities of embryos. Its main function are production of RBC, and storage of iron (in macrophages) derived from hemoglobin breakdown.  Composed of a stroma, hemopoietic cords, and sinusoidal capillaries. The stroma is fibro-reticular CT abundantly supplied by sinusoidal blood vessels lined by reticuloendothelium.  Scattered free within the reticular mesh are the bloodforming elements representing all the developmental stages of the different blood cells, making the tissue appear reddish and cellular during the 1st few years of life.  Fat cells begin to appear between the ages of 5-7 years, resulting to a change from red to the yellow variety.  At around 18 years, the red bone marrow is confined only in the regions of the vertebrae, sternum, ribs, bones of the skull and to some extent, the epiphyses of the humerus and femur.

2.Yellow or fatty bone marrow  Found in the proximal epiphyses of the humerus and femur, vertebrae, diploe of the bones of the skull, ribs and sternum of normal adults.  Composed of a fibro-reticular CT with blood vessels, nerves, and abundant fat cells crowding out the hemopoietic elements. The latter imparts a yellowish color to the tissue.  Does not play an active role in hemopoiesis, but in anemic conditions or after severe loss of blood, fatty marrow can be replaced by red marrow.

 The release of mature cells from the marrow is controlled by releasing factors, including the C3 component of the complement system; hormones (glucocorticoids and androgens); and some bacterial toxins.  In abnormal bone marrow or diseases where suppression and enhancement of proliferation of more than one type of stem cell can occur, there are reduced numbers of some cell types, e.g. aplastic anemia (decreased production of hemopoietic cells), coinciding with increased numbers of others, e.g. leukemia (abnormal proliferation of leukocytes). In myelogenous and monocytic leukemias of the bone marrow (lymphocytic leukemias occur in lymphoid tissues), the patient is usually anemic and prone to infection.  Bone marrow aspiration and the use of monoclonal antibodies specific to the membranes of precursor blood cells aid in the correct diagnosis of leukemias.

Chronic myeloid leukemia

Chronic lymphocytic leukemia

HEMOPOIETIC STEM CELLS

 Adult blood cells originate from a pluripotential hemopoietic stem cell (PHSC) in the bone marrow, the hematocytoblast.  These give rise to unipotential progenitor or precursor (blasts) cells that proliferate, acquire morphologic differentiations, and become colonyforming cells or units (CFC or CFU) with various potentialities.  These cells become lymphoid elements (lymphocytes, monocytes), and myeloid elements (RBC, granulocytes and megakaryocytes).  Growth factors, colony-stimulating factors (CSF), or hemopoietins (e.g., erythropoietins, leukopoietins) affect cell proliferation and differentiation.

 G-CSF stimulates formation of granulocytes, enhances their metabolism, and stimulates leukemic cells; GM-CSF stimulates production of granulocytes and macrophages; M-CSF stimulates formation of macrophages and increases their antitumor activity; Interleukin-3 stimulates production of all myeloid cells; EPO or erythropoietin stimulates RBC formation.  Potential therapeutic uses of growth factors include:  increasing number of blood cells in diseases or induced conditions that induce low blood counts (chemotherapy, irradiation)  increasing efficiency of bone marrow transplants  enhancing host defenses in patients with malignancies, infectious and immunodeficient diseases  enhancing treatment of parasitic diseases.

ERYTHROPOIESIS  Maintenance of the normal number of circulating RBC depend on:  Hypoxic stimulus to the marrow mediated by erythropoietin, synthesized by the renal interstitial cells  Adequate supplies of: iron for hemoglobin production, ascorbic acid, vitamin B12, intrinsic factor (present in gastric juice) which function as precursors of coenzymes in erythropoiesis.  The stages of development take only about 3 days.  Principal processes involved are reduction in size, acquisition of hemoglobin, condensation of nuclear chromatin, and eventual loss of nucleus and organelles.

1. Proerythroblast (rubriblast) earliest stage of the RBC series. It is larger than hemacytoblast; vesicular nucleus with a thin rim of heterochromatin and a distinct nucleolus; more basophilic cytoplasm than hemacytoblast due to increased RNA content. 2. Basophil erythroblast (prorubricyte) smaller than the former, nucleus with dense heterochromatin; nucleolus obscured; basophilic due to increased number of polyribosomes for hemoglobin synthesis 3. Polychromatophil erythroblast (rubricyte) stain purplish-blue to lilac due to its pink cytoplasm; nucleus has denser chromatin network and smaller in size 4. Normoblast (metarubricyte) smaller, densely basophilic nucleus; acidophilic cytoplasm due to increased hemoglobin; gradually becomes pyknotic; extrudes the rigid nucleus as it squeezes through the pores of sinusoidal walls 5. Reticulocyte same size as the normoblast but without a nucleus; less than 1% of erythrocyte count 6. Erythrocyte

Summary of erythrocyte maturation. The stippled part of the cytoplasm (on the left) shows the continuous increase in hemoglobin concentration from proerythroblast to erythrocyte. There is also a gradual decrease in nuclear volume and an increase in chromatin condensation, followed by extrusion of a pyknotic nucleus. The times are the average life span of each cell type. In the graph, 100% represents the highest recorded concentrations of hemoglobin and RNA.

GRANULOPOIESIS  Leukopoietin similar to erythropoietin has also been postulated to influence granulopoiesis.  Neutrophilia in bacterial infection or stress (e.g. intense muscular activity), increases the number of leukocytes in circulating compartment without its increased production.  The total time for a myeloblast to emerge as a mature neutrophil is about 11 days.

1.Myeloblast- finely dispersed chromatin and nucleoli; most immature serial cell 2.Promyelocyte larger than hemacytoblast; rounded or oval nucleus with dense heterochromatin and indistinct nucleoulus; basophilic cytoplasm but with localized acidophilic areas; densely scattered azurophil granules thought to represent primary lysosomes.

3.Myelocyte reduction in size and basophilia; nucleus indented to horseshoe-shape as maturation progresses; reduction of azurophil granules as specific granules characteristic of 3 granulocytes appear 4.Metamyelocyte juvenile granulocyte form are small, with characteristic granular content; band cell (stab) have more indented nucleus; as cell ages, nucleus acquires segmented nucleus and enter sinusoids 5.Granular leukocyte

 Normally, neutrophils pass through several functionally and anatomically defined compartments in their development: 1.medullary formation compartment- mitotic proliferation (3 days), and maturation (4 days) 2.medullary storage compartment (4 days)- acts as a buffer system, capable of releasing mature neutrophils on demand. 3.circulating compartment- neutrophils are suspended in plasma and circulating in blood vessels. 4.marginating compartment- neutrophils in capillaries and excluded from circulation by vasoconstriction, or adherent to lung endothelium.  There is a constant interchange of cells between the latter 2 compartments.

Functional compartments of neutrophils 1: Medullary formation compartment. 2: Medullary storage (reserve) compartment. 3: Circulating compartment. 4: Marginating compartment. The size of each compartment is roughly proportional to the number of cells.

LYMPHOPOIESIS- small lymphocytes originating from the bone marrow migrate and populate the spleen, thymus and lymph nodes. Invasion by foreign proteins stimulate undifferentiated reticular cells (lymphoblasts) within the stroma of lymphoid tissue to:  Differentiate into large and medium-sized lymphocytes, & enter circulation via the lymphatics as small lymphocytes.  Form plasma cells responsible for antibody production from lymphoblasts or from immunocompetent lymphocytes. MONOPOIESIS- monocytes originate from a pool of precursor cells in the CFU-M (monoblasts) of the bone marrow, after which they migrate into CT to become tissue macrophages.  In the marrow, immature monocytes possess a round nucleus, few ER and no ribosomes; prominent Golgi complex and precursors of azurophil granules.  Mature cells are smaller, with numerous azurophil granules which contain hydrolytic enzymes of primary lysosomes.

MEGAKARYOPOIESIS 1.Megakaryoblast large nucleus, often indented with dense peripheral heterochromatin; divides repeatedly without cytokinesis to form next stage 2.Megakaryocyte giant cells; nucleus completely lobed; cytoplasm finely granular and exhibits patchy basophilia; cell outline is indistinct, with cytoplasmic processes extending through sinusoids.  When mature, peripheral cytoplasmic processes extending from plasma membrane (demarcation membranes) become pinched off to become 4,000-8,000 platelets.  Megakaryocytes then become shrunken and their nuclei fragment.

Section of bone marrow showing various stages of megakaryocyte development (14), several adipocytes (*), and blood sinusoids (arrowheads).

Drawing showing the passage of erythrocytes, leukocytes, and platelets across a sinusoid capillary in red bone marrow. Because erythrocytes (unlike leukocytes) do not have sufficient motility to cross the wall of the sinusoid, they are believed to enter the sinusoid by a pressure gradient that exists across its wall. Leukocytes, after the action of releasing substances, cross the wall of the sinusoid by their own activity. Megakaryocytes form thin processes that cross the wall of the sinusoid and fragment at their tips, liberating the platelets.

Section of stimulated red bone marrow. Note 4 mitotic figures (arrows) and a plasma cell (arrowhead). Regions of erythropoiesis and of granulopoiesis are also evident. Most immature granulocytes are in the myelocyte stage: their cytoplasm contains large, dark-stained azurophilic granules and small, less darkly stained specific granules.

Section of red bone marrow with a group of erythropoietic cells (upper right) and a group of neutrophilopoietic cells (lower left). The immature granulocytes shown have mostly azurophilic granules in their cytoplasm and therefore are myelocytes.

Section of red bone marrow showing an immature megakaryocyte in the upper right corner. There is also a large group of erythropoietic cells (delimited by a broken line) and sparse immature neutrophils (arrowheads).

A megakaryocyte in mitosis (center) surrounded by erythropoietic cells with a mitotic figure (arrowhead). The arrow indicates an erythroblast extruding its nucleus

Section of bone marrow with an adult megakaryocyte and several granulocytes, mainly neutrophils in the myelocyte stage with many azurophilic granules and few, less darkly stained specific granules. A mitotic figure is indicated by an arrowhead.