Haematopoiesis
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Haematopoiesis. Lab 1. Bone Marrow Examination. Cellularity: (normal, decreased, increased) Evaluation of cell lines: • Erythropoiesis • Thrombopoiesis (Megakaryopoiesis) • Myelopoiesis Parasites, neoplastic cells. Bone Marrow Examination. The common indications are:

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Haematopoiesis

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Haematopoiesis

Haematopoiesis

Lab 1


Bone marrow examination

Bone Marrow Examination

  • Cellularity:(normal, decreased, increased)

  • Evaluation of cell lines:• Erythropoiesis• Thrombopoiesis (Megakaryopoiesis) • Myelopoiesis

  • Parasites, neoplastic cells


Bone marrow examination1

Bone Marrow Examination

The common indications are:

  • Persistent neutropenia

  • Persistent leukocytosis

  • Unexplained thrombocytopenia

  • Persistent thrombocytosis

  • Poorly regenerative anaemia

  • Abnormal blood cell morphology

  • Unexplained presence of immature cells in blood


Cellularity

Cellularity

The cellularity of bone marrow is estimated by examining the proportion of cells versus fat present in particles. If the particles are composed of more than 75% cells, the marrow is interpreted as hypercellular, and if the particles are composed of more than 75% fat, the marrow is interpreted as hypocellular. The overall cellularity of bone marrow decreases with age.

Histological section of canine bone marrow. Pink bony trabeculae are present in the lower left corner, lower right corner, and top, and surround the haematopoietic cells and fat. The fat is the round to oval, clear areas. The erythrocytic and granulocytic precursor cells are the many small, round purple structures. The larger, densely staining purple structures distributed throughout the marrow space are megakaryocytes.


Cellularity normal

Cellularity: Normal

Bone marrow particles filled with cells Two Megakaryocytes

If 10 to 20 Megakaryocytes are seen per field using the IOx objective, the numbers are likely increased


Cellularity decreased

Cellularity: Decreased

Hypoplasia: almost no nucleated cells, empty bm-particles


Cellularity decreased1

Cellularity: Decreased

Myelofibrosis: bm-particles containingconnective tissue, and stromal cells


Cellularity increased

Cellularity: Increased

Increased cellularity can indicate increased cell production (regenerative response) or neoplasia


Other considerations

Other considerations

  • Meyloid:Erythroid (M:E) ratio

    • Usually between 0.7 to 1.7

    • Lower ratio is usually in large animals

    • high M:E ratio could indicate the presence of either increased granulocytic cells or decreased erythroid cells

  • Stainable Iron

    • Hemosiderin appears gray to black

    • It may be seen within macrophages or as free material

    • It is decreased or absent in iron-deficient animals, and may be increased in association with all anaemias

    • Bone marrow iron increases in some animals with age


Haematopoiesis

EVALUATION OF CELL LINES: ERYTHROPOIESIS

Morphologic changes that occur as cells of the erythroid series undergo maturation include diminution in size, decrease in N:C ratio, progressive nuclear condensation, and the appearance of red cytoplasmic colour as hemoglobin is synthesized and accumulates within the cytoplasm.


Erythropoiesis

ERYTHROPOIESIS

Rubriblast. Is relatively large cells with high N:C ratio and intensely basophilic cytoplasm, resulting from the presence of many polyribosomes. The nucleus of the rubriblast is usually nearly perfectly round and the chromatin is finely granular, containing one or two pale-blue to medium-blue nucleoli.

Prorubricyte. Is a large round cell with a round nucleus with a coarsely granular chromatin pattern. This cell typically lacks a nucleolus. There is a small amount of deep blue cytoplasm with often a prominent perinuclear clear zone.

Rubricyte. is a round cell with a round, centrally located nucleus; it is smaller than the prorubricyte. The coarsely granular chromatin is more condensed compared with the earlier stages of development, and irregular clear areas are present between the chromatin clumps. Later rubricytes stain more red (contain more haemoglobin).


Haematopoiesis

ERYTHROPOIESIS


Haematopoiesis

EVALUATION OF CELL LINES: MEGAKARYOPOIESIS


Megakaryopoiesis

Megakaryopoiesis

Megakaryoblast, promegakaryocyte, and megakaryocyte

Megakaryocyte


Osteoblast osteoclast

Osteoblast / Osteoclast

An example of other cells found in a bone marrow slide and that should be differentiated from a megakaryocyte. In osteoclast nuclei are clearly separate, in contrast to the fused nuclear material present in platelet precursors.


Haematopoiesis

EVALUATION OF CELL LINES: MYELOPOIESIS

The early granulocytic precursors have oval to indented nuclei and blue cytoplasm. The larger immature forms have small, pink cytoplasmic granules. The cytoplasm becomes less blue as the cells mature.


Myelopoiesis

MYELOPOIESIS


Myelopoiesis1

MYELOPOIESIS

The myeloblast is a large, round to oval cell with a round to oval nucleus with a finely stippled chromatin pattern and usually prominent nucleolus or multiple nucleoli. There is a small to moderate amount of blue cytoplasm and no prominent cytoplasmic granules.

The promyelocyte is a large, round to oval cell with a round to oval nucleus. The nuclear chromatin pattern is finely granular. The moderate amount of cytoplasm is blue and contains multiple, pink to purple granules, which are primary granules.

The neutrophilic myelocyte is a round cell that is smaller than the myeloblast and the progranulocyte. The nucleus is round to oval and may contain a single indentation. The chromatin pattern is finely to moderately granular. Nucleoli are not present. The moderate amounts of blue cytoplasm contain multiple secondary granules, also called specific granules. These secondary granules are pink for the neutrophilic lineage and difficult to see. The secondary granules for the eosinophilic and basophilic lineages are generally reddish and purple respectively.


Myelopoiesis2

MYELOPOIESIS

The neutrophilic metamyelocyte is a round cell with a kidney-shaped nucleus. The chromatin is moderately granular and more condensed than that of the myelocyte. The moderate amounts of blue cytoplasm contain secondary granules, which are difficult to see. The secondary granules of the eosinophilic and basophilic metamyelocyte are generally reddish and purple respectively.

The band neutrophil is a round cell with a horseshoe-shaped nucleus. The cytoplasm is blue to light blue and contains secondary granules. These granules are difficult to see in the band neutrophil. The secondary granules of the band eosinophil and basophil are generally reddish and purple respectively.

The segmented neutrophil is a small round cell with a single nucleus, which has multiple segmentations. The nuclear chromatin is very condensed. There is a moderate amount of light blue to pink cytoplasm.


Erythroid or granulocyte precursors

Erythroid or granulocyte precursors!

  • As a general rule, erythroid precursors are smaller, have more nearly spherical nuclei with more condensed nuclear chromatin

  • Erythroid precursors have darker cytoplasm than do granulocyte precursors at similar maturation stages.

  • Consequently, smaller and darker cells, observed by scanning marrow smears at low power, are usually erythroid precursors (unless lymphocytes are increased in numbers), and the larger, paler cells are usually granulocyte precursors.


Other cells of the bone marrow

Other cells of the bone marrow

  • Lymphocytes

  • Monocytic series

    • (monoblast, promonocyte, and monocyte)

  • Macrophages

  • Mast cells

  • Plasma cells

  • Basket cells


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