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Bone Marrow, Clinical laboratory techniques lecture
tissue enclosed by bone (cortical and cancellous)
hematopoietic cells, adipose tissue and supportive tissue
red and yellow (all red early in life)
Red bone marrow
All is red in young animal, turns yellow with age.
Adults, can be found in proximal humerus, femur, sternum, ribs, vertebral bodies
yellow bone marrow
mainly fatty tissue, no hematopoietic function. Can turn red if needed.
precursers to cells found in blood or tissue
includes cells in mitotic (proliferation) pool and post-mitotic (maturation) pool
Includes erythrocytes, leukocytes, megakaryocytes
Things you can learn from bone marrow
Hematologic abnormalities (nonregenerative anemia, penias, persistent cytosis', abnormal blood cell morphology, unexplained presence of immature cells and regenerative or not in a horse.)
What stage cancer is in (Lymphoma, mast cell tumors)
Blood chemistry abnormalities (hyperglobulinemia, hypercalcemia)
aspirate vs. biopsy
aspirate is easier/faster/cheaper (just stain and examine)
biopsy cuts a solid core of marrow, has to be fixed, decalcified, embedded, sectioned and stained prior to examination by a pathologist. More accurate evaluation.
don't use in patients with bleeding problems (hemostatic diathesis). Could be hemorrhage.
Ribs and sternum of horse can lead to hemothorax or cardiac tampanade (pressure on heart)
Post biopsy infection
Where to perform bone marrow aspirate in dog
Trochanteric fossa of proximal femur
Where to perform bone marrow aspirate in cat
Proximal femur and proximal humerus
Where to perform bone marrow aspirate in horse, cow, camel
Ilium, ribs, sternum
Technique of bone marrow aspirate
General anesthesia or sedation, local anesthetic (both skin and periosteum)
needle cuts through muscle
Rotate needle into bone
attach syringe, pull back just until you see marrow (release quickly to avoid blood contamination)
prepare smears immediatly
drop of marrow, spread with another slide
place vertically on absorbant surface to further remedy blood contamination
air dry, stain with Romanowsky (Wright-type)
Jamshidi needle, stylette, shepherds crook
Order of erythropoeisis
prolychromatophilic erythrocyte (reticulocyte)
most immature recognizable RBC
between rubriblast and rubricyte. Cytoplasm more abundant and blue
between prorubricyte and metarubricyte. Most mature stage where mitosis can occur
most mature RBC that contains a nucleus
reticulocyte. Anucleated, blue-pink, larger than RBC
Erythroid maturation facts
nuclei gets smaller, chromatin clumps
cytoplasm goes from blue-grey to orange-red as RNA is lost
reticulocytes and RBCs migrate
Takes 3-5 days.
released from kidney in response to hypoxia. Stimulates stem cell differentiation into rubriblast. High enough concentration shortens marrow transit time and graduates early
Hormones that promote erythropoiesis
androgens, glucocorticoids, insulin, growth hormone, thyroid hormone
hormones that inhibit erythropoiesis
Polymorphonuclear leukocytes (PMNs)
Includes all three granules--neutrophils, basophils, eosinophils
Granulocyte (myeloid) maturation
Same for neutrophil, basophil and eosinophil
myelocyte (first differentiation)
earliest recognizable myeloid cell. No granules. Before promyelocyte (progranulocyte)
slightly larger than precurser myeloblast. Also larger than follower myelocyte
Nucleus is central or on one side (eccentric)
cytoplasm has primary granules.
smaller than precurser promyelocyte, larger than metamyelocyte.
primary granules replaced by secondary (specific) granules
smaller than precurser myelocyte, larger than follower band granulocyte.
kidney shaped indention in nucleus. Secondary granules.
smaller than metamyelocyte, larger than segmented granulocyte.
U-shaped or deeply indented nucleus. Secondary granules
smaller than band. Lobulated or markedly constricted nuclei, secondary granules.
Monocytes in bone marrow
difficult to differentiate because nucleus can look like anything and resembles myeloid series.
monoblast, promonocyte, monocyte in bone marrow
large multinucleated cell whose cytoplasmic fragments become platelets.
megakaryoblast, promegakaryocyte (nucleus separates), megakaryocyte.
Cells seen in bone marrow
RBC, monocytes, neutrophils
plasma cells (differentiated lymphocyte that produces hemoglobin)
lymphoblast (rare, lymphoproliferative disorders)
macrophages, osteoblasts, osteoclasts,
similar to plasma cells in appearance. In young animals and those with bone remodeling (fracture)
large multinucleated cell with nucleus separated.
normally present in very low concentration, rare in marrow.
Fibrocytes and Fibroblasts
seen infrequently because don't exfoliate easily.
Bone marrow evaluation
: whole slide for heterogenous and overall cellularity
: (25-75% cells), otherwise hyper- or hypocellular. Decreases with age. Increases with need.
: of RBC and WBC (complete, orderly, increase in # with each stage of development
Myeloid to Erythroid ratio (M:E)
: examine 500 cells, granulocytic/nRBC (hemodilution a problem)
Reticulocyte couont in horses
myeloid to erythroid ratio. normal is anywhere form 0.5:1 to 3:1. Interpret in light of CBC.
What should you see most of in bone marrow?
mature cells, due to mitosis
High M:E ratio indicates
high level of granulocytes or low level of erythrocytes
Low M:E ratio indicates
low level of granulocytes or high level of erythrocyes
reversible stem cell disorders of marrow
transient, usually stem cell recovers (if no complications)
Initial neutropenia, thrombocytopenia and nonregenerative anemia may follow if disorder lasts for 1-2 weeks.
Caused by viral (F. panleukopenia or parvo), drugs (like chemo), chemicals
irreversible stem cell injury in marrow
irreversible, cause not always understood. Can be chemicals or radiation, or FeLV.
4 types. Aplasia or hypoplasia, myelodysplasia, myeloproliferative disease, myelodysplasia becoming cancer.
4 types of irreversible stem cell injury
aplasia or hypoplasia (absent or insufficient production)
myelodysplasia (abnormal development)
myeloproliferative disease (neoplastic production)
myelodysplasia progressing into neoplasia over time.
irreversible stem cell injury with hypocellular to acellular marrow.
Presents with selective, severe and nonregenerative anemia or pancytopenia (nonregen anemia includes thrombocytopenia and neutropenia)
Red cell aplasia in dogs likely immune mediated.
abnormal development with variable manifestations of subtle, morphological changes in blood cells.
Usually includes some kind of cytopenia (neutropenia, nonregenerative anemia, thrombocytopenia)
Most common in cats, very rare in others
scarring of bone marrow, develops in response to marrow injury.
Can be caused by anything toxic to hematopoietic cells--damages microvasculature and leads to necrosis and fibrosis (dying and scarring)
myeloproliferative and lymphoproliferative disorders (leukemias)
neoplastic proliferation of hematopoietic cells within marrow.
Neoplastic cells found in peripheral blood or bone marrow.
neoplasms derived from bone marrow erythrocytes, granulocytes, monocytes and megakaryocytes. (monocytic leukemia, eosinophilic leukemia, etc.)
neoplasms from lymphocytes or plasma cells.
More immature than mature cells is bad.
ex. acute lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell myeloma.
Infiltration and replacement of marrow by blast cells that proliferate but do not mature or function in any useful way.
any hematopoietic cell line can be involved.
progression is rapid (death within days or weeks without bone marrow transplant)
some infiltration of marrow. Malignant cells mature partially and retain some function.
Affected can survive months to years with little/no treatment. Don't notice right away, much better prognosis.