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BMS247 Stem Cells > L4 - In Vitro Stem Cell Models > Flashcards

L4 - In Vitro Stem Cell Models Flashcards

1
Q

Define pluripotent cell

A

Potency to be able to generate all of the cell types of an embryo including the germ cells

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2
Q

Define multipotent cell

A

Capable of differentiation into only a few lineages

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3
Q

What are the cells called that surround the inner cell mad

A

trophectoderm

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4
Q

Give examples of pluripotent markers expressed by pluripotent cells

A

Nanog
Oct4
Sox2

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5
Q

What are the two hallmarks of pluripotency

A

Descriptive and functional

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6
Q

Descritptive test for pluripoteny

A

Expression of various pluripotency markers (Nanog, Oct4, Sox2)

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7
Q

Functional test for pluripotency

A

Grafting of pluripotent cells onto a kidney of a host mouse will give rise to a teratoma

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8
Q

What cell types are containined in a teratoma

A

All of the cell types

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9
Q

3 types of ectoderm

A

Surface
Neural
Neural crest

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10
Q

What does the ectoderm go on to form

A

Skin

Nervous system

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11
Q

4 types of mesoderm

A

Axial
APraxail
Intermediate
Lateral plate

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12
Q

What does the mesoderm go onto form

A

Blood
Heart
Muscle
Kidney

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13
Q

Whar happens to potency as devlopment proceeds

A

Becomes restricited

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14
Q

What are used to mimich the niche in

MOUSE

HUMAN

A

Leukaeamia inhibitory facotr (LIF), BMP

FGF2 and TGFB

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15
Q

Describe one method of capturing ES cells

A

Obtain the cells of the inner cell mass and dissociate
Plate on a layer of feeder cells
Once eSCs have divided a few times then disaggregate and replate

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16
Q

What factors would be used to re-program an adult somatic cell to a pluripotent fate

A

Oct4
Sox2
Klf4
C-myc

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17
Q

Describe the gene expression in iPS cells

A

Express the main pluripotency markers (Nanog, Oct4 and Sox2)

No expression of any genes which are indictaive of differentiation

18
Q

Describe some of the characteristics of iPS cells

A

Single cell able to generate many identical daughters (self-renewal)

Formation of teratoms in permissive environment

When reintroduced into an embryo they contribute to normal development

19
Q

What is a marker expressed by neural stem cells

A

RC2

20
Q

Describe a method for capturing multipotent stem cells

A

Dissociate cells

Plate on laminin in the presence of cytokines FGF2 and EGF

21
Q

Describe the 3D method for in vitro differentiation

A

Remove signals keeping cells in an undifferentiated state

Grow in aggregates (embryoid bodies/organoids)

Cell have functionality and express the right markers at the right time dependent on their fate

22
Q

Advantage of the 3D method of in vitro differentiation

A

Resembles embryonic development more closely

23
Q

Disadvantage of the 3D method of in vitro differentiation

A

Diffiuclt to observe/dissect the roles of individual signals

24
Q

Describe the 2D (Adherent) method of in vitro differentiation

A

Plate defined number of cells on the right substrate
Remove the signals keeping cells from differentiating
Grow in defined medium with appropriate levels of signals

25
Q

Advantages (2) of 2D/Abberate in vitro differentiation

A

More traceable system for live imaging

Easier to diisect roles of specific signals

26
Q

Disadvantage of 2D/Abberant in vitro differentiation

A

Don’t get same cell-cell interactions which would be seen in vivo

27
Q

What is microcephaly

A

Neurodevelopmental disorder in which infants are born with an abnormally small brain

28
Q

Why may in vitro differentiation be needed to model microcephaly

A

Mice models fail to replicate the condition

29
Q

How may microcephaly be modelling in iPS cells

A

Take a skin biopsy and culture with Oct4, Sox2, Klf4 and myc to get to the iPS cells
Formation of a cerebral organoid

30
Q

What can be seen when iPS cells are made to model microcephaly

A

fewer DCX neurones and fewer nerual progenitors

31
Q

Potential applications of microcephaly models using iPS cells

A

Small molecule screens to identify suitable drugs

32
Q

What is familiail dysautonomia

A

Genetic disorder affecting the development and survival of the neruones controlling involuntary actions

May also affect the NS in activites related to senses

33
Q

Early symptoms of familial dysautonomia

A 
Poor muscle tone 
Feeding difficulties 
Poor growth 
Lack of tears 
Frequent lung infections 
Difficulty maintaining body temperature
34
Q

Late symptoms of familial dysautonomia

A 
Delayed speech and walking 
Spinal curvature 
Reduced perception of pain/temp 
Poor growth 
UNstable BP
35
Q

familial dysautonomia usually caused by a mutation in which gene

A

IKBKAP

36
Q

Parkinsons affects how many

A

1:500

37
Q

Symtpoms of parkinsons

A 
Resting temor 
Slowness of movement 
Rigidity 
Dementia 
Anxiety
38
Q

Parkinsons characterised by the …

A

Loss of mDA neurones of the substantia nigra

39
Q

How can cell replacement be used in parkinsons

A

Generate h-ES derived mDA neurones

Transplant into mouse model

Evidence of improvement in motor function

40
Q

What may be used as a marker in Parkinsons treatment

A

TH

Tyrosin hydroxylase - enzyme involved in the synthesis of dopamine

41
Q

Cell replacement has also be used in the treatment of

A

AMD - replacing RPE cells

BMS247 Stem Cells (14 decks)

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