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clinical pathology > 4- hematopoiesis and bone marrow eval > Flashcards

4- hematopoiesis and bone marrow eval Flashcards

1
Q

In the extravascular space what is the stromal elements of the hematopoietic microenvironment

A

Stromal elements: endothelial cells, reticular (fibroblast-like) cells, adipocytes and extracellular matrix

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2
Q

in the extravascular space what are the accessory cells of the hematopoietic microenvironment

A

accessory cells: macrophages, T and B lymphocytes

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3
Q

what are the types hematopoietic cells

A
  • hematopoietic stem cells (HSC)
  • hematopoietic progenitor cells (HPC)
  • recognizable blood cell precursors
  • mature leukocyte storage pools, esp neutrophils
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4
Q

what are bone marrow stromal cells produced by

A

produced by mesenchymal stem cells

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5
Q

what do endothelial cells do in regards to bone marrow

A

they regulate the trans endothelial movement of cells btwn blood and them marrow extravascular space

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6
Q

what do reticular cells do in regards to bone marrow

A

fibroblastic-type cells they provide structural support for the bone marrow

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7
Q

what do both endothelial cells and reticular cells do

A
  • synthesis of growth factor and extracellular matrix
  • also support hematopoiesis through cell-cell contact
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8
Q

slide 9

A
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9
Q

HSCs and HPCs have what kind of appearance

A

lymphocyte appearance

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10
Q

HSCs are generally…

A

**positive for the CD34 surface antigen **

—- early HPCs are also CD34+

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11
Q

what is CD an abbreviation for

A

clusters of differentiation antigens

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12
Q

what are the properties of Hematopoietic stem cells (HSCs)

A
  • proliferation
    -sustained self replication
  • replicate slowly (once every 8-10 weeks)
  • capacity to differentiate into all blood cell types and some tissue cell types (macrophages, dendritic cells, osteoclasts, mast cells)
  • <0.0001% of nucleated cells in cat marrow
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13
Q

what do hematopoietic stem cells produce

A

common lymphoid progenitor (CLP)
— B lymphocytes, T lymphocytes, NK cells

common myeloid progenitor (CMP)
— non-lymphoid blood cells, macrophages, dendritic cells, osteoclasts, mast cells

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14
Q

what are the properties of hematopoietic progenitor cells (HPCs)

A
  • proliferation
  • limited self replication - not sustained without replenishment from HSCs
  • replicate more rapidly than HSCs
  • differentiation but with more restricted lineage potential than HSCs
  • about 1% of bone marrow cells in adults, but higher in neonates
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15
Q

what are hematopoietic growth factors (HGFs)

A

HGFs are glycoprotein cytokines that promote the proliferation, maturation, and survival of hematopoietic cells

—— HGF includes colony stimulating factors (CSFs), interleukins and poietins

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16
Q

where are hematopoietic growth factors produced

A

HGFs are produced locally in the marrow (paracrine and autocrine) and/or by cells in peripheral tissues (endocrine ex. erythropoietin from the kidney)

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17
Q

what are the early acting factors of HGFs

A

early acting factors include stem cell factor (SCF) and fIt3 ligand (FL). these factors require the presence of other cytokines

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18
Q

what are the intermediate-acting factors for HGFs

A

The intermediate-acting factors include IL3 and GM-CSF

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19
Q

what are the late acting factors for HGFs

A

late acting factors include G-CSF, M-CSF, EPO, TPO, and IL5

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20
Q

what types of indirect stimulation causes the release of HGFs

A

during inflammation, TNFa, and IL1 stimulate stromal and accessory cells to release various HGFs

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21
Q

slide 19 canine precursor cells photo

A
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22
Q

slide 20 erythropoiesis photo

A
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23
Q

what are erythroid islands

A

-erythroid cells develop around macrophages
- macrophages help create microenvironment
- phagocytize expelled nuclei and damaged or aged erythrocytes
- store iron released from phagocytized erythrocytes

add photo

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24
Q

Explain the properties of reticulocytes

A
  • formed when metarubicytes expel their nuclei, usually while still bound to macrophages
  • one rubriblast produces about 16 reticulocytes in about 4 days
  • reticulocyte maturation occurs in marrow, blood, and spleen, depending on the species
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25
Q

what does erythropoietin (EPO) do

A

—- glycoprotein growth factor
promotes proliferation, differentiation, and survival erythroid progenitor cells and early erythroid precursors (ex rubriblasts)

inhibition of apoptosis is the main mechanism of action

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26
Q

what are the sites of EPO synthesis

A

renal interstitial cells (primary site in adults)
- increased synthesis in response to tissue hypoxia in the kidney

extrarenal production
- liver, especially in the mammalian fetus
- bone marrow macrophages and erythroid cells

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27
Q

neutrophil production slide 28

A
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28
Q

what are the neutrophilic growth factors

A
  • early and intermediate acting factors

G-CSF - late acting factor
— granulocyte progenitors to myeloblasts
— increased cell division, decrease marrow transit time

Cells producing various growth factors
– fibroblasts and endothelial cells
– T lymphocytes and mononuclear phagocytes

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29
Q

slide 31

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30
Q

slide 32

A
31
Q

explain eosinophil production

A
  • marrow transit time 1 week or less
  • marrow storage pool like neutrophils
  • early and intermediate acting factors
  • IL5 from TH2 lymphocytes for terminal maturation
32
Q

how are mast cells produced

A

mast cells progenitor (MaP) released form marrow to blood

  • mast cells develop in tissues

-* stem cell factor promotes development *

33
Q

what are monocytopoiesis

A

short generation time and little marrow reserve

  • M-CSF is a late activation factor stimulation monocyte production
34
Q

monocytes may become

A
  • *macrophages *or inflammatory dendritic cells in tissues
  • osteoclasts in bone marrow
  • depends on the amounts of various cytokines presents
35
Q

what are osteoclasts

A

osteoclasts are multinucleated cells which degrade bone
- develop from monocyte fusion

36
Q

lymphopoiesis - slide 41

A
37
Q

thrombopoiesis slide 42

A
38
Q

megakaryocytes properties

A

Depending on size - 1000 to 3000 platelets produced/ megakaryocyte

— Megakaryocytes are located just outside the vascular sinuses

proplatelet processes extend into the vascular sinuses and platelets from the proplatelet processes within the vasculature

39
Q

how are non mammals different

A
  • no megakaryocytes
  • thromboblasts produce thrombocytes
  • the only giant cell in marrow is the osteoclast
40
Q

slide 47

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41
Q

slide 48

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42
Q

what promotes the stimulation of thrombopoiesis

A

early and intermediate- acting growth factors
– proliferation and expansion of megakaryocyte progenitor cells

Thrombopoietin (TPO)
- promotes differentiation of progenitor cells into megakaryocytes
- stimulates increased endomitosis, resulting in increased ploidy and size

43
Q

where is thrombopoietin produced

A

major site of production - endothelial cells of liver

– inflammatory cytokine IL6 stimulates TPO synthesis by liver during inflammation

44
Q

what is the purpose of recombinant growth factors

A

recomb human EPO and G-CSF have been used to treat animals
- they work for a short period of time, but antibodies are made against these glycoproteins, which limits their effectiveness

These antibodies may even cross- react with animals own EPO and G-CSF, when can make matters worse

dog and cat recombinant growth factors have been made, but to my knowledge are not commercially aval

45
Q

list the reasons for bone marrow exam

A
  • unexplained cytopenias in blood
  • proliferative abnormalities with unexplained left shifts, nucleated erythroid cells, or blast cells in blood
  • staging lymphomas or mast cell tumors
  • evaluate iron stores (except in cats)
  • evaluate focal lesions in bone (infections or neoplastic)
  • unexplained hyperproteinemia in blood (ex plasma cell neoplasms)
  • unexplained hypercalcemia (ex T-cell neoplasms)
46
Q

what are bone marrow aspirate smears

A

simple, fast, relatively inexpensive
– ID individual cell types and stages with routine blood stains
– prussian blue stain for iron

used for cytochemistry and immunocytochemistry to ID neoplastic cells

47
Q

How is an aspirate collected (with anticoag)

A
  • aspirate syringe contains EDTA in sterile saline solu
  • aspirate material is expelled in a petri dish and a pipette is used to collect particles to use in squash preparations
    (this allows for more slides to be prepped for special stains)
48
Q

what are bone marrow core biopsys

A

they give a better eval of overall cellularity done after unsuccessful aspirate attempts
- diagnosis of myelofibrosis
- Identification of focal lesions that may occur with metastatic neoplasia, necrosis, or granulomatous inflammation

49
Q

slide 69

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50
Q

slide 70

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51
Q

what is evaluated during a bone marrow exam

A

scan for particles and est, overall cellularity
- evaluate the number, morphology, and stages of megakaryocyte developments
- evaluate the number, morphology, and development of erythrocytic and granulocytic cells
—————-Calculate the M:E ratio (granulocytic: erythrocytic)

evaluate the number and morphology of other cell types present including lymphocytes, plasma cells, and macrophages.

assess the amount of hemosiderin present

52
Q

normal marrow cellularity on a dog

A
53
Q

slide 74

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54
Q

slide 75

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55
Q

what do you see regarding megakaryocytes in bone marrow aspirate

A

most large particles have several megakaryocytes
- normally 3-15 megakaryocytes within particles per low-power field (10x)

**Normally 80-90% are mature **

56
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slide 77

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57
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58
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59
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61
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62
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63
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64
Q

how do you interpretation of M:E ratio

A

knowledge of overall marrow cellularity, Hct and total neutrophil count in blood is critical

**- If the absolute blood neutrophil count is normal, changes in the M:E ratio generally reflect changes in the erythroid series **

**- If the Hct is normal, changes in M:E generally reflect changes in the neutrophilic series **

  • if both Hct and total neutrophil count are abnormal, the interpretation is more difficult
  • if the marrow is markedly hypoplastic of filled with neoplastic cells, the M:E ratio is of little value
65
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66
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69
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sli

70
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74
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clinical pathology (18 decks)

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