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BMS 114: Stem cells and society > Embryonic stem cells > Flashcards

Embryonic stem cells Flashcards

1
Q

What are XEN?

A

Extra embryonic stem cells

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2
Q

What are TS?

A

Trophoblast stem cells

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3
Q

What are mES and hES?

A

Mouse embryonic stem cells

Human embryonic stem cells

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4
Q

What are EpiSC?

A
  • Epiblast stem cells

- Derived just before gastrulation (post-implantation)

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5
Q

Examples of pluripotent stem cells?

A 
XEN
TS
mES
hES
EpiSC
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6
Q

What is the trophoblast?

A

Cells forming the outer layer of a blastocyst (the trophoectoderm)

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7
Q

Where have XEN and TS cells been derived from?

A

Mouse NOT the human

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8
Q

What are 8 properties of ES cells?

A

1) Derived from the inner cell mass of blastocysts

2) Non transfomed
- But high telomerase acvtivity

3) Indefinite proliferative potential
- High amplification potential

4) Stable diploid karyotype

5) Clonogenic
- Whole population can be re-derived from a single cell

6) Pluripotent
- Make all 3 germ layers

7) Carry particular markers
- Surface markers
- Genes

8) Can be incorporated into chimeras
- ONLY MOUSE ES CELLS (not human)

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9
Q

Which stem cells are naive and what does this mean?

A

Mouse ES cells

  • Don’t express lineage markers
  • Only express SC genes
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10
Q

Which stem cells are primed and what does this mean?

A

Mouse EpiSC and human PSC

  • Express lineage markers
  • Start to switch on other genes, as well as having SC genes turned on
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11
Q

How can you characterise PSC?

A

1) Surface markers
- Proteins on the surface

2) Genetically normal
- 46 XX or 46XY

3) Epigeneticically normal?
- Methylation
- Impriniting

4) Gene expression

5) Functional tests
- What can they do?
- Differentiation
- Clonogenic assays

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12
Q

How are antibodies used to characterise cells?

A
  • Cells carry different combinations of marker proteins on their surface
  • Different antibodies recognise specific proteins on the surface
  • Probe them
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13
Q

What analysis techniques are involved in picturing antibodies?

A

Immunohistochemistry (homogenate)

Immunofluorescence (in situ)

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14
Q

What controls the dose of gene expression?

A
  • Bilateral expression of genes

OR

  • Maternal imprints or paternal imprints of genes
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15
Q

How are pluripotent stem cells different from eachother?

A
  • Different characteristics depending upon where cultured from (mouse/human) and the timings
  • Characteristics are: gene expression, surface markers, epigenetic and functional (differentiation potential and single-cell re-plating ability)
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16
Q

How are human ES cells derived?

A

1) Take fertilized embryo
2) Form the blastocyt and ‘hatch’ outside of the zona pellucida

3) Remove the outer trophoectoderm layer
- By taking antibody which recognises the trophectoderm

4) Add compliment to the antibody to trigger cell lysis of the trophectoderm
5) Left with the inner cell mass and put into a medium

17
Q

How can PSCs be geneticall characterised?

A
  • Should be 46 XX or 46 XY
18
Q

How can PSCs be epigenetically characterised?

A

1) Methylation

2) X-inactivation
- In the female lines one X is inactivated

3) Imprinting

19
Q

Which gene is not expressed int the mouse or human ES cells but is expressed in mouse EpiS cells?

20
Q

What are ‘gatekeeper’ genes of pluripotency’?

A
  • Genes expressed by all ES cells (both naive and primed)
  • Required to maintain chromatin in an ‘open’ state
  • Oct 4
  • Nanog
  • Sox2
21
Q

Can all ES cells (naive or primed) from teratomas?

A

Yes

- If cells are pluripotent, they should be able to form ALL three germ layers

22
Q

What are the advantages of the embryoid body method of testing differentiation?

A
  • 3D structure
23
Q

What are the disadvantages of the embryoid body method of testing differentiation?

A
  • Not reproducible

- Not directed

24
Q

What are the advantages of the monolayer method of testing differentiation?

A

Differentiation is

  • Reproducible and
  • Directed
25
Q

What are the disadvantages of the monolayer method of testing differentiation?

A
  • No 3D structure

- Many different markers and methods

26
Q

What are the advantages of the teratoma method of testing differentiation?

A
  • 3D structure

- Extensive differentiation

27
Q

What are the disadvantages of the teratoma method of testing differentiation?

A
  • Not reproducible
  • Not directed
  • Very expensive
28
Q

What is the difference between cloning efficency of mouse and human?

A

Human cells have very poor efficency

Mouse cells have more than 80% efficency (very high)

29
Q

Where are pluripotent stem cells derived from/

A

The inner cell mass and epiblast stage embryo

30
Q

What surface markers do mice SC have that human SC don’t?

31
Q

What surface marker is shared between human and mouse SC?

32
Q

Which SC (mouse or human) has the most surface markers?

A

Human (has 7)

Mouse only has 2
SSEA1 and ALP

BMS 114: Stem cells and society (15 decks)

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