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stem cells > study session 3 > Flashcards

study session 3 Flashcards

1
Q

embroynic stem cells —– development
adult stem cells —– devellop

A

embroyonic

babies, children
adults and elderly

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2
Q

MSC different synonyms

A

Mesenchymal stem cells OR mesenchymal stromal cells OR bone marrow stem cells OR bone marrow stroma cells

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3
Q

MSC first isolated from

A

pelvic area

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4
Q

MSC cells can differentiate into

A

bone/connective tissue, but also glial, muscle hepatic cells

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5
Q

First clinical case of bone marrow stem cell

A

2008 barcelona
first tissue engeneered trachea/ bronchus
adult stem cells from bone marrow grown in large population and matured into cartilage cells (chondro). using adaptive methodology originally devised for treating osteoarth
- new chondrocytes and epithelial cells seeded into decellularised (free of donor cells) trachea seg
biosy after 1 month indicatied local bleeding meaning blood vessels had grown

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6
Q

umbillical cord vllood dervived stem cells- what markers
are they stem cells

A

neuronal, bone, adipose tissue markers
no they are non progenitor as they have immunological properties
can suppress lymphosyte prolifferation and decrease proinflammatory cytokine levels

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7
Q

umbillival cord derived stem cells used in

A

cerebral ischemia
hepatic cirrhosis
pulminary fibrosis

because they decrease inflammation

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8
Q

does umbillical cord derived stem cells have multilineage potential

A

yes

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9
Q

umbillical chord types of stem cells

A

HSC and MSC

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10
Q

pros of umbillical cord derived stem cells

A

Immune compatibility – less likely to cause immune rejection than adult stem cells (not fully developed, lack certain markers that might be/can be viewed as a foreign antigen)

Haemopoietic potential (blood disorders)

Mesenchymal potential

Less ethical issues than embryonic stem cells; ease of collection

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11
Q

what is needed to define a MSC

A

Presence of CD73, CD90 and CD105 on the cell membranes

Use of flow cytometry and fluorescent image stream analyses

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12
Q

define CD73

A

membrane-bound nucleotidase
Also known asa glycosyl phosphatidylinositol (GPI)-linked, membrane-bound glycoprotein

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13
Q

CD73 function

A

which hydrolyzes extracellular nucleoside monophosphates into bioactive nucleoside intermediates.

Surface-bound CD73 metabolizes adenosine 5′-monophosphate (AMP) to adenosine, which when released can activate one of four types of, seven transmembrane spanning adenosine receptors (which are G-protein coupled receptors) or can be internalized through dipyridamole-sensitive carriers

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14
Q

CD73 name

A

ecto-5’-nucleotidase

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15
Q

CD73 roles

A

adenosine production
regulation of immune resp
tissue repair and regeneration
endothelial function
cell migration and adhesion
cancer

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16
Q

CD73 adenosine production

A

catalyzes the conversion of extracellular ATP (adenosine triphosphate) and ADP (adenosine diphosphate) into adenosine

important in extracellular nucleotide metabolism pathway

adenosine produced is involved in various physiological and pathological process

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17
Q

CD73 and regulation of immune responses

A

inhibit T-cell proliferation,
reduce the activity of natural killer (NK) cells,
and suppress the production of pro-inflammatory cytokines.

by producing adenosine it helps modulate immune response by promoting anti inflammatory or immunosuppressive environ

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18
Q

CD73, tissue repair and regeneration

A

adenosine can contribute to tissue regeneeration and reduce damage by modulating inflammation and promoting cell surbiba;

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19
Q

CD73 cell migration and adhesion

A

effects on ECM and signalling pathways partially mediated by adensosine

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20
Q

CD73 and cancer

A

tumor microenvironment- produce adenosine promove immune invasion by tumours

heko avoid immune survaillence and promote more favourable env for growth

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21
Q

dangers of unapproved stem cell therapy _IT IS RISK

A
  • Infections: Risk of infections at the injection site or system-wide if sterility isn’t maintained.
    T - Tumor Formation: Stem cells might cause abnormal cell growth, leading to tumors.
    I - Immune Reactions: Immune responses may attack the stem cells, causing inflammation or rejection.
    S - Site Risks: Injection in delicate areas (like spine or eyes) can cause serious damage.
    R - Regulatory Gaps: Clinics without FDA approval may lack safety and quality oversight.
    I - Inconsistent Quality: Lack of standardization means varying cell quality and source safety.
    S - Side Effects: Side effects can include swelling, pain, or even worsening conditions.
    K - Knowledge Gaps: False claims and unproven therapies may mislead patients.
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22
Q

infection and stem cell

A

Procedures involving stem cell injections can lead to infections at the injection site or even systemic infections if sterility isn’t maintained. Contamination with bacteria, fungi, or other pathogens during processing can pose a serious risk.

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23
Q

immune reaction an stem cell

A

Stem cells can trigger immune responses if the body recognizes them as foreign. This can lead to inflammation, pain, or more severe complications if the immune system aggressively attacks the introduced cells.

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24
Q

tumour formation and stem cell

A

Uncontrolled cell growth is a potential risk, as some stem cells may continue to divide and form benign or malignant tumors (teratomas), especially if they haven’t been properly tested or differentiated before application.

25
Q

injection site risk

A

Local damage to tissues, nerves, or blood vessels may occur, especially when untrained personnel administer stem cell injections. For instance, injecting stem cells into sensitive areas, like the eyes or spine, carries additional risks, potentially leading to blindness or paralysis.

26
Q

lack of efficacu and false claims

A

giving patients false hope and, in some cases, deterring them from seeking effective, evidence-based treatments. This can worsen underlying conditions and lead to significant financial loss.

27
Q

regulatoryy gaps and quality control

A

Stem cell therapies are often marketed directly to consumers without proper regulatory oversight, particularly in clinics that sidestep FDA or EMA (European Medicines Agency) regulations. This lack of oversight can result in treatments with inconsistent cell quality, unknown cell sources, or improper handling.

28
Q

cell isolation from whol bood process- EASY PREP STEPS

A

EASY - Introduction to EasySep™ Direct
E - Eliminates need for red blood cell lysis and centrifugation, simplifying preparation.
A - Add Blood to a round-bottom tube; add EDTA if isolating myeloid cells.
S - Shake the RapidSpheres™ for even suspension (vortex 30 seconds).
Y - Yield Setup by adding the EasySep™ isolation cocktail to the blood.
PREP - Preparation and Initial Processing
P - Pipette and Incubate thoroughly and leave at room temperature.
R - Room Temp Incubation times vary (check product-specific info).
E - EDTA & PBS: Top-up sample with calcium- and magnesium-free PBS (plus 1 mM EDTA for myeloid cells).
P - Place in Magnet without lid; labeled cells separate from red blood cells.
STEPS - Cell Isolation and Final Collection
S - Sample Collection using a pipette to gather clear suspension.
T - Transfer and Repeat: For multiple rounds, transfer to a new tube, add RapidSpheres™ as needed.
E - Enrichment: Each separation makes the sample clearer as RBCs are depleted.
P - Pipette Carefully to avoid disturbing separated particles.
S - Support: Contact support for any protocol or product questions.

29
Q

process of cell isolation from umbillical cord, cord placenta junction and fetal placenta tissue

A

“TISSUE PREP”
TISSUE - Steps for Processing and Digestion
T - Transport and Collect: Collect umbilical cord and placenta tissues after birth, store in sterile conditions.
I - Initial Washing: Rinse tissues with saline or PBS with antibiotics to remove blood and contaminants.
S - Slice and Dissect: Cut tissue into small pieces (1-2 mm) to increase exposure for enzyme action.- scrape whartons jelly
S - Soak in Enzymes: Use collagenase or trypsin for enzymatic digestion, breaking down tissue for cell release.
U - Uniform Mixing: Agitate the tissue and enzyme mix to ensure even digestion.
E - Extract Cells: Filter the digested solution to separate cells from remaining tissue fragments.
PREP - Steps for Isolation, Culture, and Preservation
P - Pellet and Wash: Centrifuge and wash cells to concentrate and purify.
R - Resuspend in Medium: Use a growth medium to culture MSCs and ensure optimal conditions for cell growth.
E - Expand and Culture: Seed cells in plates or flasks, allowing them to adhere and proliferate.
P - Preserve or Proceed: Cryopreserve cells for future use, or proceed with characterization assays.

30
Q

adippose tissue functions

A
  • storage site lipids
    actice endocrine organ
    metabolism
    iflammation
    tissue repair
31
Q

adipose mesynvhymal stem cell names

A

Adipose-derived adult stem cells
Adipose-derived adult stromal cells
Adipose-derived stromal cells
Adipose mesenchymal cells
Adipose-derived stromal/stem cells

32
Q

what cells can be found in adipose tissue

A
  • adipocytes
    preadipocytes
    mesynchymal stem cells
    leukocytes
    fibroblasts
    endothelial cells
    smooth muscle cells
33
Q

what are preadipocytes

A

progenitor cells committed to the adipocyte lineage (not yet mature adip’s)

34
Q

how is adipose tissue obtained

A

liposuction or excision,

and stem cells are systematically isolated from samples

35
Q

adipose stem cells are derived from the

A

stronmal vascular fraction- SVF

36
Q

what is SVF

A

heterogenous mixture of cells
High proliferative and angiogenic capacity, relevant to regenerative practices

37
Q

what cells can SVF contain

A

mesynchymal stem/ stromal
endothelial progenitor
smooth muscle
M2 monocytes/ macrophages
regulatory T cells
pericytes

38
Q

how is SVF obtained

A

liposuction or excision
Mince tissue
Collagenase exposure
Filter solution through a 100um filter
Centrifuge to obtain pellet (SVF)
Remove supernatant (contains some adipocytes)
Resuspend pellet.. (further processing, i.e. cell culture, flow cytometry, differentiation)

39
Q

how is SVF manufactures

A

solid fat- mincing, collegenase, inactivation
digesyed fat liquid- RBC lysis, washes, centrifuge, filter
svf at bottom- aquous llayer- fatty layer top

40
Q

what CD cells should adipose derived stemcells be positive for

A

73
90
105

41
Q

what CD cells should adipose derived stemcells be NEGATIVE for

A

45

42
Q

characteristics of adipose derived stem ce;;s

A

plastic adhesion
self renew
abundant source
multi lineage capacity
low immunogenicity

43
Q

why is CD45 negative in adipose

A

CD45 (protein tyrosine phosphatase receptor type C): This protein is a transmembrane protein that is expressed on hematopoietic cells, including leukocytes and lymphocytes. It is involved in the regulation of immune cell function and is typically absent from mesenchymal stem cells.

44
Q

differentiation od adipose derived stem cells

A

adipocytes
chondro
ostep
hepato
endothelial
beta islets
muscle
keratino
neuronal lineages
glial

45
Q

clinical applications of adipose derived stem cells

A

wound healing
breast augmentation
cosmetology
hair regeneration
antiaging
muscular dystrophy
bone regeneration

46
Q

human endometrium 2 layer

A

functional
basal

47
Q

functional layer endometrium

A

always undergos reorganisation

48
Q

basal layer endometrium

A

mostly loose conjunctive tissue

49
Q

stem cells of endometrium obtained through

A

Hysterectomy

Collection of menstrual blood (in vessels containing heparin and antibiotics)
Easy adhesion to plastic

50
Q

menstrual derived stem cells benifits

A

multipotent and non invasive

51
Q

menstrual dervived mesynchymal stem cells can become

A

Cardiomyocytes
Respiratory epithelium
Neural cells
Myocytes
Endothelial cells
Pancreatic cells
Osteocytes
Hepatocytes
Adipocytes

52
Q

perinatal tissue

A

placenta
amniotic fluid
umbilical cord
whartons jelly

53
Q

what is whartons jelly

A

mucopolysaccharide
mucous connective tissue containing some fibroblasts and macrophages (insulates/protects umbilical cord)

54
Q

perinatal tissue is what stem cell

A

MSC and HSC

55
Q

IPSC

A

induced pluripotent stem cells

56
Q

what are ipcs

A

somatic cells reprogramed to revert to embroyonic like pluripotent stage

through ectopic expression of TF

adipocyte stem cells can generate ipsc

57
Q

creatubg iPS cells

A

isolatep- skin or fibroblast- grow in a dish
treat cells with reprogramming factors
wait a few weeks
pluripotent stem cells
change culture conditions to stimulate differentiate in viariety cells
- blood, gut, cardiac

58
Q

advantages of iPSC

A

Ethical considerations
Personalized medicine (patient’s own tissue)
Unlimited supply (adult stem cell population decreases)
Disease modelling (patient’s own genetic mutations/genotype)
Pluripotent (not just multipotent)
Lower risk of tumor formation (than embryonic stem cells)
Ease of collection

59
Q

applications iPSC

A

Fractured bones, tissue injuries
Joint/cartilage tissue in rheumatoid arthritis
Wound healing (MSCs – growth factors, angiogenesis, etc.) – used in diabetes for chronic ulcers/sores, burns
Immunomodulation (anti-inflammatory, graft vs host disease, autoimmune)
Clinical trials ongoing for myocardial infarction
Neurodegeneration (as well as spinal cord injuries)
Vehicles for drug delivery (affinity towards sites of injury, cancer/tumours)
Research models (lab vs clinic)
Eye conditions

stem cells (4 decks)

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