Bone marrow
Prof. Dr. Malak A. Al-yawerObjectives
Identify the major site of hematopoiesis in the fetus and normal adult. state the types of bone marrow outline the compartments of red bone marrow describe the types and distinctive characteristic of stem cells know the differences between stem cells, progenitor cells, blast cells and mature cells Review the stem cell theory of hematopoiesis. Discuss growth factors involved in the stimulation and regulation of hematopoietic activity. Review maturation stages of each cell lineage, including changes that occur as cells mature.Bone marrow
Is a soft connective tissue occupies the medullary cavity of long bones and all the spaces between the trabeculae of spongy bone It accounts for approximately 5% of the body weight in humansTwo varieties of marrow are recognized Red marrow Yellow marrow
Red marrowIt is the only site for genesis of blood cells in adults Is the only type found in fetal and young bones but in adult it is restricted to the vertebrae , sternum , ribs , cranial bones and epiphysis of long bones
Amount of red marrow varies with the age
Childhood- red marrow is 100% of bone marrow and present in virtually every bone. Adults- red marrow is 50% of bone marrow presenting in sternum, ribs, pelvis and skull. 70 years- red marrow reduced to 30% of the bone marrow.Yellow marrow
Consists in main of fat cells which have gradually replaced the other marrow elements . With an adequate stimulus , yellow marrow may resume the character of red marrow and play an active role in the process of blood development .
Red bone marrow has
A vascular compartmentAn extravascular compartment
A femoral head with a cortex of cortical bone and medulla of trabecular bone. Both red bone marrow and a focus of yellow bone marrow are visible.Vascular compartment
Is supplied by a nutrient artery which branches into central longitudinal arteries which send out radial branches that eventually open into sinuses. These sinuses converge into a central vein that carries the blood out of the bone marrow into the general circulation The marrow does not have lymphatic drainageExtravascular compartment
Is composed of : Stroma of reticular connective tissue hematopoietic cords sinusoidal capillariesStroma is composed of
reticular tissue 1. reticular cells 2.Reticular fibers macrophages and adipose cells MatrixReticular cells
Stellate in shape and are in contact with each other along extended cellular processes.
Function of reticular cells: Act as a meshwork to support and protect the haemopoietic cells Formation of reticular fibers Phagocytosis They transfer to adipocytes by accumalating lipid in their contents
Matrix
Contain: collagen type I , III laminin , fibronectin and proteoglycanLaminin, fibronectin, and another cell-binding substance, hemonectin, interact with cell receptors to bind cells to the stroma.
sinusoidal capillaries
The sinusoids are formed by a discontinuous layer of endothelial cells. An external discontinuous layer of reticular cells and a loose net of reticular fibers reinforce the sinusoidal capillaries.Bone marrow barrier
The blood vessels of the bone marrow constitute a barrier, inhibiting immature blood cells from leaving the marrow. Only mature blood cells contain the membrane proteins required to attach to and pass the blood vessel endothelium. Hematopoietic stem cells may also cross the bone marrow barrier, and may thus be harvested from blood.Stem cells
are pluripotential cells capable of self-renewal. Some of their daughter cells form specific, irreversibly differentiated cell types, and other daughter cells remain stem cells.Stem cells
They retain the ability to renew themselves through mitotic cell division and can differentiate into a diverse range of specialized cell types. They found in all multi-cellular organism .Two broad types of mammalian stem cells
1. Embryonic stem cells that are found in blastocysts 2. Adult stem cells that are found in adult tissuesEmbryonic Stem Cells
are derived from the inner cell mass of the embryo. Because these cells are pluripotent and can virtually form any cell or tissue type, they have the potential for curing a variety of diseases, including diabetes, Alzheimer and Parkinson diseases, anemias, spinal cord injuries, and many others.ES cells may be obtained
reproductive cloning. therapeutic cloning (somatic nuclear transfer)Reproductive cloning
ES cells may be obtained from embryos after in vitro fertilizationThis approach has the disadvantage that the cells may cause immune rejection, since they would not be genetically identical to their hosts. Another issue with this approach is based on ethical considerations, since the cells are derived from fertilized viable embryos.
Therapeutic cloning (somatic nuclear transfer)
take nuclei from adult cells (e.g., skin) and introduce them into enucleated oocytes. Oocytes are stimulated to differentiate into blastocysts, and ES cells are harvested.Since the cells are derived from the host, they are compatible genetically and since fertilization is not involved, the technique is less controversial.
Adult Stem Cells
Adult tissues contain stem cells that also may prove valuable in treating diseases. These cells are restricted in their ability to form different cell types and, therefore, are multipotent, not pluripotent.Adult stem cells isolated from rat brains have been used to cure Parkinson disease in rats, suggesting that the approach has promise. Disadvantages of the approach include the slow rates of cell division characteristic of these cells and their scarcity, which makes them difficult to isolate in sufficient numbers for experiments.
Hematopoiesis
takes place in the extravascular compartment The currently accepted theory on how this process works is called the monophyletic theory which simply means that a single type of stem cells gives rise to all mature blood cells in the body. This stem cells is called the pluripotential stem cells.pleuripotential stem cell
It is believed that all blood cells arise from a single type of stem cell in the bone marrow . It is called a pleuripotential stem cell because it can porduce all blood cell types .Pleuripotential stem cell
proliferate and form 1. lymphoid multipotential cells: one cell lineage that will become lymphocytes . And 2. myeloid multipotential cells : another lineage that will form the myeloid cells ( granulocytes, monocytes, erythrocytes and megakaryocytes)SITES OF HEMATOPOIESIS
Site of hematopoiesisAge
yolk sac then liver
Embryo
Spleen
3rd to 7th month
marrow cavity - esp. granulocytes and platelets
4th and 5th months
marrow cavity - erythrocytes
7th month
mostly bone marrow; spleen and liver when needed
Birth
number of active sites in bone marrow decreases but retain ability for hematopoiesis
Birth to maturity
bone marrow of skull, ribs, sternum, vertebral column, pelvis, proximal ends of femurs
Adult
Hematopoiesis depends on
favorable microenvironmental conditions and the presence of growth factors.
The microenvironmental conditions are furnished by cells of the stroma of hematopoietic organs, which produce an adequate extracellular matrix.
A general view of hematopoiesis shows that
both the potential for differentiation and the self-renewing capacity of the initial cells gradually decrease. the mitotic response to growth factors gradually increases, attaining its maximum in the middle of the process. From that point on, mitotic activity decreases, morphological characteristics and functional activity develop, and mature cells are formedstem cells: This cell can produce all blood cell types Low mitotic activity Self renewing Scarce in the bone marrow Cannot be morphologically distinguished ( resemble large lymphocyte )
Progenitor cells : Could be unipotential or bipotential High mitotic activity Self renewing Common in marrow and lymphoid organs Cannot be morphologically distinguished ( resemble large lymphocyte )
Precursor cells( blast ): Monopotential cells High mitotic activity Not self renewing Common in marrow and lymphoid organs Beginning of morphologic differentiation
Mature cells : No mitotic activity Abundant in the blood and haematopoietic organs Clear morphologic differentiation
Hematopoiesis is a compartmentalized process within the hematopoietic tissue
erythropoiesis taking place in distinct anatomical units (erythroblastic islands); granulopoiesis occurs in less distinct foci megakaryopoiesis occurs adjacent to the sinus endothelium.Upon maturation, the hematopoietic cells, regulated by the reticular cells, traverse the wall of the venous sinuses to enter the bloodstream
Leukocytes, after the action of releasing substances, cross the wall of the sinusoid by their own activity. 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. Megakaryocytes form thin processes (proplatelet processes) that cross the wall of the sinusoid and fragment at their tips, liberating the platelets.
Maturation of erythrocytes
Mainly regulated by erythropoietin released by the kidneys; also influenced by androgens
Maturation of erythrocytes
1. pluripotential cell2. Myeloid multipotential cell3. Erythrocyte – colony forming cell 4. Proerythroblast (pronormoblasts) : large cell , rounded nucleus coarse chromatin , visible nucleoli, intense basophilia of the cytoplasmMaturation of erythrocytes
5. Basophilic erythroblast (basophilic normoblasts) : condensed nucleus , no visible nucleoli , strongly basophilic cytoplasm because of free ribosomes and polyribosomes. 6. Polychromatophilic erythroblast (polychromatophilic normoblasts) : mixed color cytoplasm purplish blue to greyMaturation of erythrocytes
7. Orthochromatophilic erythroblast : the amount of haemoglobin is the same as that of erythrocyte . Nucleus with dense and compact chromatin ---------pyknotic --------- extruded from the cell with a thin rim of cytoplasm and plasma membrane8. Reticulocyte: youngest erythrocyte containing a delicate reticulum the clumped ribosomes responsible for the distinctive staining of the reticulocytes are degraded within 24 hours 9. Erythrocyte : anucleated and biconcave in peripheral blood
Several major changes take place during maturation of erythrocyte
1. cell volume decreases 2. nucleoli diminish in size until they become invisible 3. nuclear diameter decrease and chromatin increase until the nucleus become pyknotic and extruded from the cell 4. gradual decrease in the number of polyribosomes ( basophilia )with a simultaneous increase in the amount of haemoglobin( acidophilic protein ) 5. mitochondria and other organelles gradually disapearElectron micrograph of red bone marrow
. Four erythroblasts in successive stages of maturation are seen (E1, E2, E3, and E4). As the cell matures, its chromatin becomes gradually condensed, the accumulation of hemoglobin increases the electron density of the cytoplasm, and the mitochondria (M) decrease in number.Granulopiosis
Regulated by GM-CSFGranulopiosis
1. pluripotential stem cells 2. myeloid multipotential stem cells 3. granulocyte colony forming cell 4. Myeloblast : ovoid nucleus with 2 or more nucleoli , basophilic cytoplasm (abundance of RER and ribosomes ) 5. Promyelocyte : round or oval nucleus , occasionally indented , basophilic cytoplasm containing azurophilic granules ( lysosomal enzyme and myeloperoxidase )Granulopiosis
6. Myelocyte : appearance of specific granules and the developing myelocytes can be distinguished into 3 types : Neutrophilic myelocyte Acidophilic myelocyte Basophilic myelocyteGranulopiosis
7. Metamyelocyte : nuclei irregular in shape known as band form , cytoplasm with increasing free ribosomes , mitochondria and RER . Neutrophilic metamyelocyte Acidophilic metamyelocyte Basophilic metamyelocyteGranulopiosis
8. Mature granulocyte : Neutrophils : horse –shoe or S- shaped nucleus Acidophil : nucleus with 2 lobes , common in connective tissue of certain organs ( intestine epithelium ) Basophil : nucleus is large , irregularAzurophilic granules
1st appear in promyelocyte Derived from the inner cisternae of Golgi complex Decrease in number with frequent division and maturation They are lysosomes , it’s histochemical structure myeloperoxidase and acid phosphataseSpecific granules
Initial appearance in myelocyte Derived from an outer cisternae of Golgi complex Increase in number with maturation neutrophil contain alkaline phosphatase and antibacterial lysozyme Acidophil contain sulphatase , peroxidase and histaminase Basophil contain heparin and histamineDrawing illustrating the sequence of gene expression in the maturation of granulocytes. Azurophilic granules are blue; specific granules are pink.
MEDICAL APPLICATION
The appearance of large numbers of immature neutrophils (band cells) in the blood is called a shift to the left and is clinically significant, usually indicating bacterial infection.Monopoiesis
Maturation of monocytes Is regulated by GM-CSFMaturation of monocytes
1. Pluripotential cell 2. Myeloid multipotential cell 3. Monocyte colony – forming cell4. monoblasts 5. Promonocyte : large cell 18 micrometer in diameter , slightly indented nucleus with lacy chromatin and evident nucleoli , basophilic cytoplasm6. Monocyte : indented nucleus cytoplasm contain large amount of RER, extensive Golgi complex and granule condensation These granules are primary lysosomes, which are observed as fine azurophilic granules in blood monocytes.
Monocytes migrate into the circulation where they remain for about 8 hours before migrating into the connective tissue in the connective tissue they increase in size, acquire multiple lysosomes and become active in phagocytosis (macrophages) life span of macrophages in different tissues may be up to several months
Lymphopoiesis
Maturation of lymphocyteMaturation of lymphocyte
1. Pluripotential cell 2. Lymphoid multipotential cells : migrate to lymphoid organs 3. Lymphocyte colony forming cell 4. Lymphoblast : large cells , large nucleus with prominent nucleoli , basophilic cytoplasm capable of incorporating [3H]thymidine and dividing two or three times to form prolymphocytes
Maturation of lymphocyte
5. Prolymphocyte : smaller with condensed chromatin , nucleoli less obvious , few azurophilic granules appear in the cytoplasm , no cell surface receptor that mark them as T or B lymphocyteMaturation of lymphocyte
6. B and T lymphocytes : in the thymus or bone marrow , these cells synthesize cell surface receptors but they are not recognized as distinct cell types using routine histological procedure . They can be recognized by immunohistochemistryMedical application
Leukemias are malignant clones of leukocyte precursors. They occur in lymphoid tissue (lymphocytic leukemias) bone marrow (myelogenous and monocytic leukemias). In these diseases, there is usually a release of large numbers of immature cells into the blood. The symptoms of leukemias are a consequence of this shift in cell proliferation, with a lack of some cell types and excessive production of others (which are often abnormal in function). The patient is usually anemic and prone to infection.Thrombopoiesis
Maturation of platelets Regulated by thrombopoietin (TPO) Mainly produced by the liverMaturation of platelets
1. Pluripotential cell 2. Myeloid multipotential cell 3. Megakaryocyte forming cell 4. Megakaryoblast : large cell ( 15-50 micrometer ) , large ovoid or kidney shaped nucleus with numerous nucleoli ( DNA 30X as much as a normal cell ) , cytoplasm is homogenous and basophilicMaturation of platelets
5. Megakaryocyte : giant cell ( 35-150 micrometer), irregular lobulated nucleus , coarse chromatin , no visible nucleoli , Cytoplasm contain numerous mitochondria RER, extensive Golgi complex , conspicuous granules contain biologically active substances such as platelet derived growth factor , fibroblast growth factor The demarkation membranes arise from numerous invaginations of the plasma membrane through out the cytoplasmThe demarkation Membranes
Electron micrographs indicate that megakaryocytic cytoplasm can fragment through the formation of tiny channels arising from rows of vesicle The vesicles fuse with their neighbors and establish continuity with the cell membrane, producing an extensive system of tubular platelet demarcation channels hat subdivide the cytoplasm into hundreds of platelets each with its covering
Electron micrograph of a megakaryocyte showing a lobulated nucleus (N) and numerous cytoplasmic granules. The demarcation membranes are visible as tubular profiles.
Cells of the megakaryocyte series shown in a bone marrow smear. Note the formation of platelets at the lower end of the megakaryocyte.
Section of bone marrow showing various stages of megakaryocyte development (1—4), several adipocytes (*), and blood sinusoids (arrowheads).
MEDICAL APPLICATION
In certain forms of thrombocytopenic purpura, a disease in which the number of blood platelets is reduced, the platelets appear to be bound to the cytoplasm of the megakaryocytes, indicating a defect in the liberation mechanism of these corpuscles. The life span of platelets is approximately 10 days.Clinical evaluation
Clinical evaluation Tests of bone marrow functionSmall amounts of myeloid tissue is removed from bone cavity and examined under microscope. Bone marrow aspiration- cytology and maturation Bone marrow biopsy- cellularity and architecture
Bone marrow aspiration and biopsy
Bone marrow transplant (or hematopoietic stem cell transplantation)
is a very complicated and risky process, and therefore applied only to patients with life-threatening diseases (who are resistant to chemotherapy). Mostly patients with congenital neutropenia, aplastic anemia, sickle-cell disease, neuroblastoma, lymphoma and many other diseases are treated by stem cell / bone marrow transplantation.