13 August 2017

Lecture #2: The Hematopoetic System



Hematopoeisis or hemopoeisis is the presence of blood cells production by the blood forming organs.

Erythropoeisis – process of producing red blood cells
Leukopoeisis – process of producing white blood cells
Myelopoeisis – process of producing granulocytes
Lymphopoeisis – process of producing lymphocytes
Thrombopoeisis – process of producing thrombocytes or blood platelets

The hematopoietic system is highly specialized for both the production of new blood cells and the removal of those which are old and worn out. To  keep the total number of cells fairly constant, new ones must be added to the blood each day to replace the old ones which are removed.

Undifferentiated cells of the mesenchyme and endothelial cells of the primitive blood spaces give rise to blood cells in the embryo. Later, in due course, production of blood cells becomes organoid and the liver and spleen become the main sites. Eventually, the bone marrow takes over. At birth, the marrow is active and red and contains only minimal amounts of fat. Later in childhood, only the proximal portions of the long bones and the flat bones are the sites of blood formation. The rest of the marrow space is occupied by fat, which can be replaced by hemopoeitic cells when the need arises.

Hematopoietic tissues – organs or tissues in which blood cell production occurs are known as hematopoietic tissues. Based on the organs and tissues involved, hematopoiesis may be called:

1. Medullary hematopoiesis

This involves the formation of normal red blood cells in the marrow. It is limited to the vertebrae, ribs, sternum, pelvis, scapulae, skull and extreme portions of humeri and femora. In infants and children, active hematopoiesis takes place in more distal portions of the extremities than in adults. Hematopoiesis occurs within the parenchyma of the bone marrow bounded by sinusoids.

The red bone marrow – produces red cells, platelets, neutrophils, eosinophils and basophils and serves as one of the sites of formation of lymphocytes, monocytes and plasmacytes.



The yellow bone marrow – is filled with fat. This fat occupies reserve space and readily goes into solution and disappears as the blood cells increases, thus making it possible for regeneration to occur on demand without interfering with vital structures.

2. Extramedullary hematopoiesis

This involves the formation of normal red blood cells outside the confines of the bone marrow. The spleen is the most commonly encountered site, but hematopoiesis in liver, lymph nodes and in the islands of the lymphatic tissue and less commonly adrenal glands, cartilage, broad ligament, thrombi, adipose tissue, intrathoracic tissue, kidneys and endosteum. These hematopoietic areas may be composed of pure or mixed erythrocytic, leukocytic or megakaryocytic tissue.

Lymphatic system – the lymphocytes are formed in the most part in the lymph nodes and in the island of lymphatic tissue located in the spleen, bone marrow, thymus, liver and intestinal tract and to a lesser extent in practically over other organ except the brain and spinal cord.

Spleen – it aids in the formation of lymphocytes and monocytes and serves as a reservoir for cells which can be delivered from the pulp into the circulation on demand.

Stomach – secretes the intrinsic factor, the presence of which is essential in the absorption of Vitamin B12 from the intestinal tract. Vitamin B12 is a requirement for the normal maturation of erythrocytes.


THEORIES OF BLOOD CELL DEVELOPMENT


There are two broad theories or schools of thought regarding the development of cells:

1. Unitarian theory states that all blood cells are derived from a hemohistioblast (reticuloendothelial stem cell) which is capable of giving rise to several types of cells.

2. Polyphyletic theory states that the first recognizable and most primitive blood cells are already differentiated and their future development along one line is already determined.

In any case, all cells arise from the stem cell.

In man, all blood cells are derived from a pluripotential stem cell. A stem cell possesses 2 different properties:

a. Property of proliferating by mitosis

b. Property of continuing to differentiate, i.e., to mature along a cell line, during which process the various more mature forms retains only a limited proliferative capacity. Pluripotential means that this stem cell has the ability to differentiate into a number of possible cell series. In other words, it may evolve in the direction of megakaryocytes or in the direction of monocytes. Although a unipotential cell can divide and thus ensure the continuation of this type of cell; it can also continue to differentiate along a specific line of cells. A myeloblast, for instance, will only give rise to granulocytes, a proerythroblast only to erythrocytes. These stem cells are also known as “committed stem cells” (pluripotential stem cell) to change into a “committed stem cell” are imperfectly understood.


MATURATION OF BLOOD CELLS


Principle of normal cell maturation

In any series of cell development, an almost infinite gradation of cells exists between the most immature “blast” form and the mature definitive cells. The mature cell is identified and characterized by certain specific features which develop during the course of maturation. These features gradually appear as the cell matures so that each nucleus must evolve gradually and appear at some stage of maturation. Since this is a normal and orderly process, blood cells exhibit constant features which lead to the description of typical stages. Since a cell is composed of several components or structures, each of these parts undergoes individual transformation. Normally, these transformations or changes occur simultaneously (synchronism) which simplifies description. Abnormal cell maturation may be characterized by different rates of maturation of the different parts (asynchronism) producing cells that are “atypical” or bizarre but whose external structure is easily analyzed as that of normal cell.


There are three changes that always occur in the transformation from an immature to a mature cell:


1. Cytoplasmic changes

a. Loss of basophilia – the cytoplasm of an immature cell is usually blue or basophilic due to its ribonucleic acid (RNA) content. The more mature the cell, the less basophilic because of the less RNA content.

b. Cytoplasmic granules – in myeloid cells, the cytoplasm is studded with granules. These granules contain some enzymes which distinguish the myeloid stem cells from other cells. Those which have affinity to the red acidic dye are called acidophilic or eosinophilic granulocytes. Those which have affinity to the blue basic dye are called basophilic granulocytes and those that have affinity to both the acid and basic dyes are called neutrophilic granulocytes.

c. Elaboration of hemoglobin – this is a special feature of the maturation of erythrocytic cells. At first, the immature cell contains no hemoglobin. Gradually, the hemoglobin starts to appear as the cell becomes mature until the most mature cell contains a standard and maximal amount of it. At the mature stage the nucleus is apparently no longer necessary and is eliminated by nucleolysis or extrusion.

2.     Nuclear changes

a. Structure and cytochemistry – the immature nucleus is round or oval and the nuclear chromatin is very delicate. As the cell matures, the chromatin strands become increasingly coarse and clumped and there is also a reduction in the number of nucleoli.

b. Shape changes – as the cell mature, the shape of the nucleus changes too. This is especially true with granulocytes in which the nucleus divides into segments or lobes as it grows older. The older the cell, the more segments or lobes the nucleus has.

3. Reduction of cell size – it is a feature of all cells except megakaryocytic series in which the immature cell is smaller than the fully developed megakaryocyte. Generally speaking, a mature cell is smaller than an immature one.


Principle of abnormal cell maturation


Pathologic hematopoiesis results in abnormal nuclear maturation, abnormal cytoplasmic differentiation and abnormal size. Their development may also be asynchronous.

1. Abnormal cytoplasmic differentiation – in erythrocytes, this is characterized by persistent cytoplasmic basophilia and late hemoglobinization. Abnormal cytoplasmic inclusion bodies may be found in the cytoplasm of both erythrocytes and leukocytes, especially in the granulocytes.

2. Abnormal nuclear maturation – in leukemia and other severe disturbances, two nuclei may be present; one may be diploid and other polyploid. The nucleus may be hypersegmented or hyposegmented.

3. Abnormal size – abnormally large cells are frequently seen in benign or malignant proliferation. In erythrocytic series, the megaloblasts are larger than normal and are even present in the mature erythrocyte. Likewise, abnormally small cells may be found.


STEM CELLS


In postnatal life in humans, erythrocytes, granulocytes, monocytes and platelets are normally produced in bone marrow. Lymphocytes are produced in the secondary lymphoid organs, as well as in the bone marrow and thymus gland.

Most bone marrow cells are morphologically recognizable precursors of granulocytes, erythrocytes or platelets. A fraction of cells in adult marrow is identifiable only as lymphocytes. Smaller number of monocytes, macrophages, endothelial cells and plasma cells are noted.

Stem cells can both reproduce themselves and give rise to more differentiated cells; they are not morphologically identifiable as stem cells. A pluripotential or multipotential stem cell is present in the marrow and gives rise to two major progenitors, the lymphoid stem cells and myeloid or hematopoietic stem cell. The latter is a common precursor cells for granulocytes and monocytes, erythrocyte and megakaryocytes.

Committed progenitor cells (committed stem cells) are characterized by their ability to form colonies in vitro in response to a soluble factor.

It is believed that the pluripotential hematopoietic stem cell is induced by certain microenvironmental influences to become the committed erythroid progenitor cell.

The committed progenitor cell for neutrophils and monocytes divides and gives rise to the myeloblast, the earliest recognizable granulocyte–monocyte precursors, under stimulation by a hormone, colony– stimulating factor for granulocytes and monocytes.

Monocytes share the same committed progenitor cell as neutrophils, the GM – CFC.

Platelets originate from polyploid megakaryocytes, the largest of all hematopoietic cells, which number less than 1% of the total nucleated marrow cells. They arise from the multipotential hematopoietic stem cells, probably directly from a committed progenitor cells.

According to current concepts, during fetal life, lymphocyte precursors originate in the bone marrow and are influenced or programmed to perform a certain function by one of the primary lymphoid organs, either the thymus gland for T–lymphocytes (T–cells) or the “bursal equivalent” for B– lymphocytes (B–cells)


NORMOBLASTIC MATURATION



RUBRIBLAST

Synonyms:      Proerythroblast or Pronormoblast
Size:                 14 – 19 micrometers
Nucleus:          round or oval; thin membrane; central or slightly eccentric
Chromatin:     fine to coarse; scanty and indistinct
Nucleoli:         1 – 2; usually pink
Cytoplasm:     small, moderately basophilic, homogenous and opaque
N/C ratio:        8:1

PRORUBRICYTE

Synonyms:      Early erythroblast
                        Basophilic normoblast
                        Basophilic erythroblast
Size:                 10 – 15 micrometers
Nucleus:          smaller than pronormoblast, round and slightly eccentric with thin membrane
Chromatin:     irregular and coarse; sparse but distinct
Nucleoli:         0 – 1
Cytoplasm:     apparently abundant; royal blue and opaque, with varying amounts of
                        hemoglobin; with Reddish tinge
N/C ratio:        6:1

RUBRICYTE

Synonyms:      Polychromatophilic erythroblast
                        Polychromatophilic normoblast
                        Intermediate erythroblast
                        Intermediate normoblast
Size:                 8 – 12 micrometers
Nucleus:          round, smaller than prorubricyte; eccentric; thick membrane
Chromatin:     coarse and clumped; irregularly condensed; distinct parachromatin
Nucleoli:         no longer visible
Cytoplasm:     relatively more abundant; varying mixture of red and blue to diffusely lilac
N/C:                 4:1

METARUBRICYTE

Synonyms:      Orthochromatic erythroblast
                        Orthochromatic normoblast
                        Late erythroblast
                        Late normoblast
Size:                 7 – 10 micrometers
Nucleus:          small, shrunken, solid black; degenerated
Chromatin:     non–linear clumped and condensed
Nucleoli:         none
Cytoplasm:     red with minimal amounts of residual blue

RETICULOCYTE

Synonyms:      Diffusely basophilic erythrocyte
                        Polychromatophilic erythrocyte
                        Vital granulated erythrocyte
                        Vital staining erythrocyte
                        Pro–erythrocyte
 Size:                8 – 10 micrometers
Nucleus:          none, instead, after supravital stain center of cell shows network of fibrillae and
                        dots or Reticulum of bluish tint (residue of RNA)
Chromatin:     None
Nucleoli:         None
Cytoplasm:     pink to reddish brown

ERYTHROCYTE

Synonyms:      Normocyte, akaryocyte, red blood cell, erythroplastid
Size:                 6 – 8 micrometers
Nucleus:          None
Chromatin:     None
Nucleoli:         None
Cytoplasm:     Pink, darker in periphery than in the center


MATURATION OF GRANULOCYTE


MYELOBLAST

Size:                 10 – 20 micrometers
Nucleus:          Round or oval, thin nuclear membrane; stippled finely reticulated; light purple
Chromatin:     abundant chromatin, pale blue or pink parachromatin
Nucleoli:         2 – 5 round or oval; pale blue
Cytoplasm:     sparse, deeply basophilic; no granules
N/C ratio:        7:1

PROMYELOCYTE

Synonyms:      Progranulocyte
Size:                 14 – 20 micrometers
Nucleus:          large, round or oval, thin membrane
Chromatin:     slightly clumping
Nucleoli:         1 – 3; pale blue; round or oval
Cytoplasm:     sparse; basophilic, contains slight purplish granules
N/C ratio:        5:1

MYELOCYTE

Size:                 10 – 19 micrometers
Nucleus:          round or oval
Chromatin:     Coarse condensed chromatin with blue or pink parachromatin
Nucleoli:         Rarely more than one; usually not visible
Cytoplasm:     moderate amount; large granules – purplish blue at early stage; granules turn
                        eosinophilic or basophilic or neutrophilic in the late or more mature stage.
N/C ratio:        2:1

METAMYELOCYTE

Synonym:        Juvenile
Size:                 10 – 28 micrometers
Nucleus:          Indented and kidney – shaped
Chromatin:     Deep purple; coarse and more clumped; scanty parachromatin
N/C ratio:        1:5:1
Nucleoli:         Not visible
Cytoplasm:     Fairly abundant; pink with eosinophilic or basophilic or neutrophilic granules

BAND

Synonyms:      Stab, Staff cell
Size:                 10 – 15 micrometers
Nucleus:          Sausage shaped; band shaped or shaped like letters C,S,Z; curved rod
Chromatin:     coarse chromatin; deep purplish blue
Nucleoli:         Not visible
Cytoplasm:     Abundant; pale blue or pink with neutrophilic, eosinophilic or basophilic granules
N/C ratio:        1:2

SEGMENTED GRANULOCYTE

Synonym:        Polymorphonuclear neutrophil, basophil or eosinophil
Size:                 10 – 15 micrometers
Chromatin:     Coarse, dense, deep purplish blue
Nucleus:          Two or more lobes or segments connected by filaments
Nucleoli:         Not visible
Cytoplasm:     Abundant; blue or pink with characteristic eosinophilic [red, neutrophilic (lilac) or
                        Basophilic (dark blue or blackish) granules
N/C ratio:        1:3

Eosinophils rarely have more than 2 segments; usually nucleus lies over the granules are coarse and red. Basophils usually have indented nucleus, rarely more than 2 segments, usually the granules are found lying over the nucleus; granules are coarse and dark blue.
Neutrophils have nuclei with more than 2 segments as much as 5. Granules are lilac and finer.


MATURATION OF LYMPHOBLAST


LYMPHOBLAST

Size:                 10 – 18 micrometers
Nucleus:          Centrally located; definite membrane, round or oval
Chromatin:     Thin strands or light red purple chromatin
                        Light blue sharply demarcated parachromatin
Nucleoli:         1 – 2 small pale blue
Cytoplasm:     Homogenous and moderately basophilic; sparse with no granule
N/C ratio:        7:5

PROLYMPHOCYTE

Size:                 10 – 18 micrometers
Nucleus:          Round or oval; slightly indented
Chromatin:     Reddish – purple densed chromatin; parachromatin not well defined
Nucleoli:         One, round, blue and well outlined
Cytoplasm:     Moderately abundant; pale blue to medium dark blue
N/C ratio:        5:1

LYMPHOCYTE

Size:                 Small lymphocyte – 6 – 8 micrometers
                        Large lymphocyte – 8 – 18 micrometers
Nucleus:          Round or oval, slightly or deeply indented, eccentric, heavy membrane, closely
                        knit
Chromatin:     Large coarse clumps of chromatin blending into sparse pale blue to pink
                        parachromatin
Nucleoli:         Generally none, occasionally present
Cytoplasm:     Clear, homogenous Robin’s egg blue or sky blue with occasional azurophilic
                        granules
N/C ratio:        5:2

                                               
MATURATION OF MONOBLAST


MONOBLAST

Size:                 12 – 18 micrometers
Nucleus:          pale staining and indented; fine strands of chromatin, abundant parachromatin
Nucleoli:         Usually none; occasionally one
Cytoplasm:     Gray blue; opaque with numerous fine dustlike lilac granules
N/C ratio:        6:1

PROMONOCYTE

Size:                 14 – 18 micrometers
Nucleus:          Moderately indented; thin membrane
Chromatin:     Fine, thread – like chromatin; abundant parachromatin
Nucleoli:         0 – 1
Cytoplasm:     Opaque, gray blue with very fine lilac granules
N/C ratio:        5:1

MONOCYTE

Size:                 12 – 18 micrometers
Nucleus:          Indented or folder over; delicate pale staining; kidney shaped; with brain – like
                        Convolutions; sprawling with blunt pseudopods
Chromatin:     Fine strands; abundant and distinct parachromatin
Nucleoli:         Occasionally blue; usually none
Cytoplasm:     Abundant; slightly gray ground glass appearance
N/C ratio:        4:1


MATURATION OF MEGAKARYOBLAST


MEGAKARYOBLAST

Size:                 25 – 35 micrometers
Nucleus:          Large oval or round
Chromatin:     Delicate purple chromatin; sparse parachromatin
Nucleoli:         2 – 6; small and indistinct
Cytoplasm:     Scanty; irregular; blue non–granular with blunt pseudopods with various shades
                        of Blue; may contain azurophilic granules
N/C ratio:        10:1

PROMEGAKARYOCYTE

Size:                 25 – 60 micrometers
Nucleus:          Irregularly large; single but may appear lobulated or multilobulated
Nucleoli:         2 – 6 small indistinct
Cytoplasm:     Basophilic; prominent blue to reddish purple granules and marginal bubbly
                        cytoplasm
N/C ratio:        6:1

MEGAKARYOCYTE

Size:                 40 – 150 micrometers
Nucleus:          multiform resembling staghorn calculi; irregular bizarre
Chromatin:     Coarse; irregularly clumped
Nucleoli:         Usually not visible
Cytoplasm:     Abundant, pale with fine blue granules and showing pseudopod – like
                        projections
N/C ratio:        1:2

The granular megakaryocyte is characterized by spreading of the red pink granules diffusely through most of the cytoplasm and further increase and spreading of nuclear lobes. In the mature megakaryocyte, the nucleus is more compact, basophilia has disappeared, and the granules are clustered into small aggregates. At an ultrastructural level, this is produced by proliferation in invaginated surface membrane (demarcating membrane) that separates the cytoplasm into individual platelets. Platelets are ultimately shed as cytoplasmic fragments by fusion of demarcation membranes. In the marrow, megakaryocytes are adjacent to sinus walls, and platelets are released into the lumen.

THROMBOCYTE

Synonym:        Blood platelet
Size:                 2 – 5 micrometers
Nucleus:          None
Nucleoli:         None
Cytoplasm:     Light blue with granules at the center (granulomere, chromomere), marginal
                        zone – Hyalomere


MATURATION OF PLASMABLAST


PLASMABLAST

Size:                 15 – 25 micrometers
Nucleus:          Round or oval; eccentric
Chromatin:     Coarse and reticulated; moderate amount and distinct parachromatin
Nucleoli:         2 – 4
Cytoplasm:     Fairly abundant, moderately or deeply basophilic with no granules
N/C ratio:        1:2

PROPLASMACYTE

Size:                 15 – 25 micrometers
Nucleus:          Oval or rough; eccentric; moderately coarse
Chromatin:     Intermediate
Nucleoli:         1 – 2; very large when abnormal
Cytoplasm:     Brilliant blue; opaque
N/C ratio:        1:2

PLASMACYTE

Synonym:        Plasma cell
Size:                 10 – 20 micrometers
Nucleus:          Round or oval; eccentric, dense and concentrated in the periphery creating the
                        so called “cart–wheel” or “spokes of a wheel” arrangement
Nucleoli:         None
Cytoplasm:     Dark blue; ovoid and somewhat fibrillary with pale clear perinuclear zone;
non–granular but may contain secretory globules called “Russell bodies” which 
may be colorless, red, pink, blue or green in color. If globules fill the cytoplasm, cell is called “grape or berry or morula” cell. The intracytoplasmic deposition or amorphous material gives rise to Mott cell or Flame cell

N/C ratio:        1:2



No comments: