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Lecture 1-18

1
Q

Stage 3 trials

A

Prolonged

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2
Q

5 vector based approaches

A

Transient transfection Adenovirus Floxed vectors Transposon mRNA

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3
Q

Which methods may be used to identify iPSCs

A

Epigenetics Protein and gene expression Differentiation Morphology

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4
Q

What cell types does the myeloid progenitor give rise to

A

Neutrophils Basophils Eosinophils Monocytes Erythrocytes

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5
Q

Name some of the functional tests for PSCs

A

Teratoma form Embroid bodies Single cell replating Spin EB and growth factors

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6
Q

Stage 1 trials

A

Safety

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7
Q

Mouse of Human SSSEA1 SSEA3 SSEA4 TRA-1-60 GCTM2 Thy1 MHC ALP

A

SSSEA1 M SSEA3 H SSEA4 H TRA-1-60 H GCTM2 H Thy1 H MHC H ALP Neither

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8
Q

DEVELOPMENTAL POTENTIAL MOUSE ES CELLS TERATOMA CHIMERA

A

TERATOMA Y CHIMERA Y

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9
Q

In the mouse embryo what is the fate of the inner cells

A

Form the epiblast then the embryo

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10
Q

What is IRES used for

A

Knock in Sequence when in transcript ribosome can bind so two different proteins are produced

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11
Q

As the primitivie streak extends what becomes more anterior

A

Node

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12
Q

What is the point of no return in the cell cycle

A

Restriction point

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13
Q

When is cyclin E found

A

G1/S

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14
Q

Chemical bases approaches

A

VPA 5-AzaC

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15
Q

Describe gastrulation

A

Epiblast cells migrate through the primitve streak emerging and endoderm and mesoderm this produces the 3 germ layers

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16
Q

Advantages of floxed based delivery

A

Efficient Can be removed

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17
Q

What are tissue specific SCs also known as

A

Adult

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18
Q

As the embryo grows what are pushed more anteriorly

A

AVE AND DVE

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19
Q

Characterstics of apoptosis

A

No loss of integrity Aggregation of chromatin at the nuclear membrane Shrinkage of the cytoplasm and nucleus condenses Mitochondria releases death signals

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20
Q

Describe the progression of AMD

A

RPE cells toward the rear of the retina malfunction and die Photoreceptors adjacent ro RPE die and malformation of choroid physically displaces the overlying neural retina

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21
Q

What can be said about the colonogenic capacity of hES cells

A

Low

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22
Q

How would you specify DA neurons

A

Supress mesoderm and promote ectoderm Inhibit BMP and TGFB

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23
Q

Describe the cell cycle of PSCs

A

G1 shorter Self renewal maintained in G2/S

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24
Q

How are embroid bodies used

A

Induce differentiation and examine the 3D structure

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25
Q

Mutations in BMP TGF-B can cause

A

Cancer

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26
Q

What virus can transduce both dividing and non dividing

A

Can transduce both dividing and non-dividing cells

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27
Q

Disadvantages of adenovirus based delivery

A

Repeated infection required for certain cell types Delayed kinetics of reprogramming

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28
Q

How can you tell RPE cells in suspension

A

Pigmented

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29
Q

Implications of the ASC model

A

Cells flow through in unidirectional manner Differentiated cells become restricted Ordered and irreversible decisions

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30
Q

Descirbe the restriction point

A

CDK4/Cyclin D –> Phsophrylation of E2F and pRb CDK2/Cyclin E –> diphosphorylation of E2F and pRb Dissociation E2f enters the nucelus

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31
Q

Stages for drug disocvery

A

Target identification Lead selection Lead optimise Pre clinical development Clinical trials phase 1 2 3

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32
Q

Give of an example of a stem cell with a medium turnover

A

Red blood cells Cardiomyocytes

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33
Q

Describe physiological assay

A

Low throughput difficult assay physiological

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34
Q

Which AMD variant is more common

A

Wet 90%

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35
Q

RESPONSE TO FACTORS MOUSE EPISC LIF NODAL/ACTIVIN FGF2

A

LIF N NODAL/ACTIVIN Y FGF2 Y

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36
Q

What happens at the metaphase-anaphase transition

A

Check whether all chromosomes attached to the spindle

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37
Q

Where in each signalling pathway may feedback act

A

Every step

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38
Q

Which two progenitors are produced by the mPSCs in the blood

A

Lymphoid progenitor and the myeloid progenitor

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39
Q

Stage 2 trails

A

Efficacy

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40
Q

INK4 family

A

Inhibit CDK4

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41
Q

CELL SURFACE MARKERS MOUSE EPISC SSEA1 SSEA3/SSEA4 ALKALINE PHOSPHATASE

A

SSEA1 Y SSEA3/SSEA4 N ALKALINE PHOSPHATASE N

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42
Q

Symptoms of AMD

A

Trouble disconcerting colours Blurred vision Decrease in visual acuity Slow recovery after exposure to bright lights

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43
Q

What is found between the RPE cells and the choroid

A

Bruchs membrane

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44
Q

Steps for induction of pluripotency

A

Somatic cells differentiated Viral transduction Colony formation IPS cells

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45
Q

Where does TFIID bind

A

The TATA box

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46
Q

Descibe activation of an RTP

A

Autophosphorylate tyrosine residues

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47
Q

Summarise ES cells

A

From embryo High prolif Highly potent High availability

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48
Q

Which types of viral transduction could be used to treat AMD

A

OCT4 SOX2 KLF4 C-MYC LIN28 NANOG

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49
Q

How would you specify ectoderm

A

Block TGF-B and BMP signalling

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50
Q

What method of gene editting may be used to bolt on other proteins

A

CAS9 defficient nuclease

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51
Q

Describe a normally functioning basal ganglia

A

DA produced by cells in the SN which affect other areas of the dorsal striatum

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52
Q

How can cultured PRE cells from post mortem material be used to treat AMD

A

In vitro human RPE retain sight with transplant into rat Can be tested with rodent head tracking

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53
Q

How can we stimulate sensory regeneration in the human ear

A

Need to remove inhibition of the stem cells

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54
Q

What is a TE

A

Transposable element which can change place in the genome

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55
Q

KIP family

A

CDK inhibitor protein

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56
Q

Describe the JAK STAT pathway

A

Cytokine receptors activate the JAK tyrosine kinase, phosphorylate themselves and other receptors STAT gene reg proteins Activated STATS dimerise and migrate to the nucleus

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57
Q

Give examples where human cell der cells lines are useful, which situations may they not be useful in

A

Useful in cancer BUT not if you wanted to study the very early stages of tumourigenesis

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58
Q

Functions of normal RPE cells

A

Regeneration of bleaches opsins Transepithelial transport Absorption of light Protection from photooxidation Secretion of growth factors Phagocytosis

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59
Q

3 germ layers

A

Endo/Ecto/Mesoderm

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60
Q

What virus can only tranduce dividing cells

A

Retrovirus

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61
Q

How can mRNA used to deliver

A

Exogenous RNA causes an ancient antiviral response causing degredation Need drugs to supress this and 5’ protected mRNA

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62
Q

By what movement do the cells move through the primitve streak

A

Involution

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63
Q

How may cell replacement be used to treat parkinsons

A

Human foetal transplant (cells in the human foetus which make and release dopamine) Cells that make DA (can be similar or not (RPE cells)

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64
Q

What can be said about RA gradients during somitogenesis

A

Highest in the middle tails down either side

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65
Q

What is the effect of TGF-B and BMP signalling

A

They exert proliferative signals to cells

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66
Q

How are proteins degraded

A

Tagged with ubiquitin Enters proteasome and degraded

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67
Q

DEVELOPMENTAL POTENTIAL HUMAN ES CELLS TERATOMA CHIMERA

A

TERATOMA Y CHIMERA N/D

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68
Q

What project radially

A

Lysine rich N terminus tails of core histones

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69
Q

Broad classes of ways in which hearing loss can be prevented

A

Anti oxidants Anti apoptopic drugs NMDA blockers Growth factors

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70
Q

How can teratoma form be a test for PSCs

A

Should form all 3 of the germ layers

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71
Q

What is the frequency range of human speech

A

2-7 kHz

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72
Q

Advantages of transposon based delivery

A

Efficient Mark ffree excision

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73
Q

3 fundamental properties of stem cells

A

Potency Self renewal Differentiation

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74
Q

Side effects of DA agonists

A

Similiar to LDOPA side effects, can also cause hallucinations

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75
Q

What do the absoptive progenitors give rise to

A

Enterocytes

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76
Q

Describe the pathway of ASCs in the muscles

A

Satellite cell Myoblast Myocyte Myotube

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77
Q

Advantages of mRNA based delivery

A

No viral componenets No genomic integration

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78
Q

What determines the cell type

A

The repitoire of genes switched on

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79
Q

Advantages to virus based delivery

A

Efficient delivery Should be silenced

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80
Q

What is special about cyclin D

A

Direct link between the EC environment and the cell cycle Target of many growth factors

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81
Q

4 signalling centres in embryos

A

Post epiblast Anterior visceral endoderm Exterior embyonic endoderm Node

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82
Q

Issues with Talens/ZFN

A

Protein:DNA interactions Hard to design

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83
Q

Disadvantages of floxed based delivery

A

Genomic integration: risk of insertional mutagenesis Expresion often maintained in iPSCs Leaves a mark

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84
Q

What happens when hair cells are damaged in chicks

A

Transdifferentiation of supporting cells Proliferative regeneration

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85
Q

What happens in G2 phase

A

Cell checks fidelity of the DNA and preparation for nuclear division

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86
Q

Disadvantages of virus based delivery

A

Genomic integration: risk of insertional mutagenesis Expression often maintained in iPSCs: increased risk of tumours

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87
Q

CELL SURFACE MARKERS HUMAN ES CELLS SSEA1 SSEA3/SSEA4 ALKALINE PHOSPHATASE

A

SSEA1 N SSEA3/SSEA4 Y ALKALINE PHOSPHATASE Y

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88
Q

In ectodermal patterning what determines the AP axis What drives neural differentiation

A

FGFs BMP

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89
Q

What is 2A used for

A

Cleaves protein and is inserted

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90
Q

What is special about the +4 LRCs

A

Can revert back to the LGR 5+

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91
Q

What do the myo-SC give rise to

A

Myoepithelium

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92
Q

What can be seen in parkinsons patients

A

Lewy bodies made of alpha synuclein

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93
Q

Define neuralation

A

FOlding of the neural plate to give the nerural tube

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94
Q

Why is atoh1 not expressed in the supporting cells

A

Intracellular domain of notch is cleaved off in the supporting cells and enters the nucelus allowing the Hes to repress Atoh1

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95
Q

Route to specifying midbrain DA neuroens

A

BMP inhib WNT SHH and Low WNT

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96
Q

Describe the fovea

A

Closely packed cones Accurate vision

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97
Q

Stages for IVF/Embryo transfer

A

Synchronise and stimulate follicle development Stimulate pre ovulatory follicle development Retrive oocytes Ova under oil and transfer to sperm solution (37 deg and 5% CO2) Capasitate sperm and ISC if necessary Check zygote is fertilised Incubate to 4 cell stage or blastocyst stage PGD Transfer 1-2 embryos back into the mother

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98
Q

Which medications may be used to treat parkinsons

A

Dopemaine agonists MAO/COMT inhibitors L-DOPA

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99
Q

General ASC model

A

Stem cell gives rise to progenitors Progenitors expand Hardwired hierarchy Asymm division ASC divide few times to protect themselves from muations

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100
Q

Purposes of using drugs to treat disease

A

Reverse disease Prevent

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101
Q

Causes of hearing loss

A

Noise Genetics Ageing disease

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102
Q

What does CRSIPR use to target correct DNA sequence

A

Uses guide RNA

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103
Q

EXCEPTION TO THE MODEL Flow isnt always unidirectional

A

Loss of basal stem cells Clara cells de differentiate to replace

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104
Q

What cell types do the lymphoid progenitor give rise to

A

NK cells B and T lymphocytes

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105
Q

Give of an example of a stem cell with a lifetime turnover

A

Lens Oocytes

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106
Q

Descirbe the niche triangle

A

Cell:soluble factors Cell:ECM environment Cell:Cell

107
Q

STEM CELLS GENES MOUSE ES CELLS OCT4 NANOG SOX2 KIF4 DPPA3 REX1 GBX2 FGF5

A

OCT4 Y NANOG Y SOX2 Y KIF4 Y DPPA3 Y REX1 Y GBX2 Y FGF5 NO

108
Q

What SCs would come from the blastocyst

A

mESC hESC EpiSC Naive human SC

109
Q

Describe the macula

A

Functional centre of the retina responsible for 20/20 vision

110
Q

At which 3 places does histone acetylation occur

A

Aetylation of lysines Phosphorylation of serine 10 in H3 Mono/di/tri methylation of lysine and arginine in core histone N terminus tails

111
Q

Describe the process for deriving the ES cells

A

Hatched blastocyst Antitrophectoderm and complement Leaves innner cell mass

112
Q

What do stem cell properties allow

A

Regeneration Development - building embryos and tissue

113
Q

What do the Lgr 5+ CBC give rise to

A

Non cycling precurrsors +4 LRC and absorptive prgenitor

114
Q

Motor symtpoms

A

Slowness of movement Decreased ability to perform spontaneous movements Postural changes Shuffling gait Resting tremor

115
Q

Which viruses may be used to induce pluripotency

A

OCT4 SOX2 KLF4 C-MYC LIN28 NANOG

116
Q

When active what does TGF act

A

SMAD 2 and 3

117
Q

Malfunction of apoptosis can lead to

A

Neurodgen disease Ischameic stroke Autoimmune disease HIV Cancer

118
Q

Describe the niche in muscles

A

Between muscle fibres and myofibres

119
Q

What tissues from the endoderm

A

Pancreas Liver

120
Q

What were the two ‘senses’ for the origin of the stem cell

A

It is the unicellular origin of life The fertellised oocyte is the origin of all cells

121
Q

When active what does BMP act

A

SMAD 1 5 8

122
Q

What else do transcription activators bind to

A

The mediator complex

123
Q

What can be said about PSCs when growing in vitro

A

Will proliferate indefinetly

124
Q

Describre high throughput screening

A

High througput Simple Not single cell so would need a target or reported

125
Q

RESPONSE TO FACTORS HUMAN ES CELLS LIF NODAL/ACTIVIN FGF2

A

LIF N NODAL/ACTIVIN Y FGF2 Y

126
Q

What does a primed cell mean

A

Expresses lineage markers

127
Q

What does the haematopoetic stem cell give rise to

A

Multipotent stem cells

128
Q

Properties of ES cells

A

Indef prolif pot Carry particular markers Pluripotent Not transformed but high telomerase Derv from inner cell mass of blastocyst Clonergenic Stable diploid karyotype

129
Q

Why would iPSCs be used

A

Capture a diseased genotype OF A HUMAN POP OR DISEASED GENOYPE

130
Q

Cyclin E pairs

131
Q

What type of signalling is used between hair cells and the supporting cells

132
Q

Give situations where cells may need to die by apoptosis

A

Cells with DNA damage Loss of endometrium Cells of the immune system - after cell mediated IR Development and morphogenesis Cancer cells Viral infected cells

133
Q

Effects of alternate splicing

A

One gene can create multiple proteins

134
Q

How does RNAi work

A

Physically blocks translation

135
Q

Positives of using PSCs to treat Parkinsons

A

Could be gorwn in large batches and immune matched

136
Q

Summarise tissue specific stem cells

A

Low prolif From adult and juvenile and foetal organs Multipotent Hard to access and isolate

137
Q

What occurs with dry AMD

A

Little no haemorhaging

138
Q

What component parts of the BG

A

Nucelus accumbens Substantia nigra striatum Globus palidus Thalamus STN

139
Q

Analysis methods of disease models

A

Antibody staining Cell counts Reporters

140
Q

Descirbe the basal ganglia in Parkinsons patients

A

SN inputs are diminished Harder to generate transient inhibition from caudate and putamen Result is tonic inhibition from GP to thalamus (unable to send inpulses to the motor cortex)

141
Q

Disadvantages of transposon based delivery

A

Clones need to be checked for excision and rearrangements

142
Q

Overall effect of acetylation of histones

A

Opens up the chromatin

143
Q

STEM CELL GENES MOUSE EPISC OCT4 NANOG SOX2 KIF4 DPPA3 REX1 GBX2 FGF5

A

OCT4 Y NANOG Y SOX2 Y KIF4 N DPPA3 N REX1 N GBX2 N FGF5 Y

144
Q

Descirbe some basic niche concepts

A

Occupancy - puts SCs near the proliferative signals Fate - signal from niche keeps them in the SC cell fate Asym div - physical organisation of the niche causes polarisation of the cells to ensure asymmetrical division

145
Q

Give of an example of a stem cell with a quick turnover

A

Most epithelia Colon crypt cells

146
Q

Problems with cell transplantation to treat AMD

A

Neural retinal restored to reveal RPE cell layer and transplanted onto the macula

147
Q

Describe WNT signalling in the ON state

A

WNT binds fizzled LRP5/6 binding complex activates dischevelled which prevents destruction of B-catenin enters nucleus

148
Q

Components and reasons for components in a human artificial chromosome

A

Telomere - maint integrity Rep origin - so can be replicated Centromere - segregation of chromosomes

149
Q

Describe the extrinsic pathway of apoptosis

A

Death ligands Death receptors Initiator caspase 8 Effector caspase 3

150
Q

What gene is expressed in the hair cells

151
Q

What part of the brain is effected in Parkinsons patients

A

Basal ganglia

152
Q

In the epiblast what does BMP induce

A

Induces lefty1

153
Q

Describe the action of nitric oxide

A

Chem ubstable Small and uncharged Cytoplasmic receptor

154
Q

Mouse EpiSC

A

From pregastrualtion emb Primed

155
Q

Disadvantages of mRNA based delivery

A

Multiple rounds of transfection Lower levels of expression than when integrated

156
Q

Two categories of parkinsons symtpoms

A

Motor dysfunction and non motor symptoms

157
Q

Define a niche

A

Defined anatomical location promoting self renewal

158
Q

Problems with cell transplantation to treat AMD

A

Restoration of central vision at the expense of area of peripheral vision Complicated operation Follow up surgery to remove compression medium

159
Q

In chick what is present during gastrulation

A

Hensons node

160
Q

What do the secretory progenitors give rise to

A

Goblet cell Enteroendocrine cell Paneth cell

161
Q

What tissues from the mesoderm

A

Heart Vascular Blood

162
Q

Describe the sets for creating a disease model

A

Wild type iPSCs –> wt cells –> wt pheno Wt iPSCS –> Dis iPSCS –> dis cells –> dis phenotype Wt iPSCS –> Dis iPSCS –> dis cells –> TREAT see effect of phenotype

163
Q

RESPONSE TO FACTORS MOUSE ES CELLS LIF NODAL/ACTIVIN FGF2

A

LIF Y NODAL/ACTIVIN N FGF2 N

164
Q

How do dopamine agaonists work

A

Not metabolised into DA but mimic the action

165
Q

What does transposase catalyse

A

Movement of the transposable element to another part of the genome

166
Q

General principle of eukaryotic transcription

A

Transcription factors bind to enhancers Allows RNA polymerase to bind to promoter region

167
Q

Process for induction of pluripotency

A

Start with somatic cells that are diff Colony form iPSC cells signal to ES cells by markers

168
Q

What activates TGF-B

A

Nodal Activin-A TGF-B

169
Q

What are the choices for stem cells

A

Differentiation Death Self renewal

170
Q

How can ASC assist in repair

A

Stop themsevles aging Prevent mutations Supply large numbers of differentiated cells

171
Q

Two conditions of using PSCs to treat Parkinsons

A

Would need to ensure differentiaed to the correct cell type Check physiologically mature

172
Q

Protein based approches

A

TAT-fusion

173
Q

Describe the niche found in the Distal tip cell in the C.elegans

A

DTC triggers self renewal in the germ line stem cells as cells move away from the DTCs they divide by meiosis forming the gametes

174
Q

Summarise iPSC properties

A

Pluripotent High prolif capacity Easily available From somatic cells

175
Q

Disadvantages of VPA 5-AzaC

A

Very inefficient

176
Q

Disadvantages of TAT fusion

A

Inefficient

177
Q

What do the Lum SC cells give rise to, finish the pathway

A

Alveolar progenitor –> Alveolar cell Ductal progenitor –> Ductal luminal cell

178
Q

TGF and BMP compete for

179
Q

Where is WNT AND TGF-B signalling restricted to

A

Post epiblast

180
Q

What can be said about the efficiency of cloning

A

Decreases with each stage and in more complex organisms

181
Q

Which enzyme is involved in siRNA

182
Q

What is used to fuse the myotubes

A

Horse serum

183
Q

Describe the valley model

A

Cellular states like rolling a ball down a hill Valleys act as barriers

184
Q

Describe high contrast screening

A

Single cell Mid throughput Diffiucult assay

185
Q

What happens at the G2 checkpoint

A

Is all DNA replicated Is the cell big enough Is environment favourable

186
Q

What forms at the highest concentration of NODAL

187
Q

Two sources of PSCS

A

Reprogrammed From the early mammalian embryo

188
Q

How can a cell transplant to treat AMD work

A

Autologous RPE transplant maitains/restores sight in humans affected

189
Q

Problems of L-dopa treatment

A

Prolonged can caused dyskineasias

190
Q

What tissues from the ectoderm

A

Non neural Neural

191
Q

What happens if you remove a SC from the niche

A

Differentiation

192
Q

Three types of elecrtical implants used what does each of them bypass

A

Auditory nerve inplant - bypasses the cochlear Brainstem implant - bypasses cochlear and auditory nerve Cochlear implant - bypasses the hair cells

193
Q

In the mouse embryo what is the fate of the outer cells

A

Extraembryonic (placenta)

194
Q

Describe WNT signalling in the OFF state

A

B-catenin associated with E-cadherin Targetted phosphorylation –> ubiquitination –> degredation of B-catenin

195
Q

What happens in G0

A

Cell withdraw from the cell cycle, replication machinery is dismantled and cell enters a quiescent state

196
Q

When is cyclin B found

197
Q

Cyclin A pairs

198
Q

What is drunsen

A

ECM build up between Bruchs membrane and RPE cells of the eyes

199
Q

What is the solution for AMD treatment

A

RPE cells in solution at high density + bare patch can be used to create an RPE cell coated patch

200
Q

What did Weismann propose in 1892

A

Germ plasm continuity

201
Q

MASC can give rise to (mammary gland)

A

Myo-SC Lum-SC

202
Q

Describe PI3 kinase activation

A

Leads to activation of Akt

203
Q

Problems with cultured RPE cells being transplanted from post mortem material to treat AMD

A

RPE cells show limitted proliferative capacity in vitro Some karyotypically abndormal RPE phenotypes degenerate with time in culture REP phenotype tends to degenerate in culture

204
Q

Why may CRISPR be preferred

A

RNA:DNA interactions Easier to design

205
Q

Functions of dopamine

A

Related to mood Stimulates smooth and balance muscle movement

206
Q

Steps for cloning

A

Take an adult somatic cell and remove nucleus Insert nucleus into an unfertilized oocyte Electric shock to stimulate uptake of the nucleus

207
Q

Possible concepts of using stem cells to treat hearing loss

A

Can we differentiate ES cells to hair cells Transplanting Awakaken endogenous stem cells Do any stem cells exist in the inner ear

208
Q

When is cyclin D found

209
Q

Advantages of VPA 5-AzaC

A

Very easy to use Cheap

210
Q

How would you specify primitive streak cells

A

TFG-B and WNT

211
Q

Describe the notch signalling pathway

A

Delta and notch both membrane bound When bind cleaves the EC domain of notch leacing the adam tace Gamma-secretase cleaves the IC domain of notch and it is able to enter the nucleus

212
Q

More Shh in the brain gives a

A

More ventral cell fate

213
Q

During cloning what happens

A

SOMATIC GENES OFF EMBRYO GENES ON DNA methylation back to the totipotent configuration Cell nucleus manipulated to reset the state

214
Q

Advantages of adenovirus based delivery

A

Lower frequency of genomic integration

215
Q

Advantages of using human PSCs

A

Differentiation to any cell fate Grow indefinetly Normal primary line

216
Q

HES

217
Q

How does L-dopa work

A

DA precurrsor which is able to cross the blood brain barrier and be metabolised to DA

218
Q

EXCEPTION TO THE MODEL Not all SCs are multipotent

A

Epidermal basal cells ONLY make karatinocytes

219
Q

Steroid hormones

A

Structurally similar to cholesterol Hydrophobic IC receptor

220
Q

Hearing loss prevalence 17-30 31-40 41-50 51-60 61-70 71-80

A

1.8 2.8 8.2 18.9 36.8 60.2

221
Q

EXCEPTION TO THE MODEL Not all cells are quiescent

A

E.g. in intestinal crypt cells SC are continuouskly dividing

222
Q

Why is age a problem with AMD tratement

A

Aged bruchs membrane, RPE cells may fail to attach

223
Q

At what stage are apical and basolateral domains established

A

Early morula

224
Q

Mouse ES

A

From blastocyst Naive

225
Q

What activates BMP

A

GDF3 Noggin

226
Q

Characteristics of cancer

A

Loss of restriction point Normal cell cycle regulatory machinery lost Loss of contact inhibition Failure to respond to growth factors Cells cant arrest

227
Q

Problems with using mouse cells lines

A

Physical dif Physiological dif Genetic dif SOme mutations in the BLM genes are fatal in mice but cause disease in humans

228
Q

Characteristics of necrosis

A

Total cell lysis No vesicle formation Loss of integrity Swelling of the cytoplasm Disintegration of organelles

229
Q

What occurs with wet AMD

A

Growth of new blood vessels within the retinal tissue and subsequent haemorrhage

230
Q

Effects of DNA methylation

A

Directly blocks TFIID binding Recruitment of histone de-acetylases

231
Q

What can Ras activate, describe this activation

A

MAP kinases M kkk M kk M k Gene expression/regulatory activity

232
Q

What is the ASC in the small intestine crypt villus unit

A

Lgr 5+ CBC

233
Q

How could PSCs be characterised by genetics

A

Imprininting X inactivation Methylation

234
Q

Prevalence in 40+ 65+ 80+

A

0.4% 1% 10%

235
Q

DEVELOPMENTAL POTENTIAL MOUSE EPISC TERATOMA CHIMERA

A

TERATOMA Y CHIMERA N

236
Q

What are the 3 main types of non-integrating delivery of reprogramming factors

A

Vector based Protein based Chemical bases

237
Q

Stratergies of generating disease models

A

Healthy PSCs then gene edit Grow indefinetly Somatic cells –>> iPSCs and then gene edit

238
Q

How may surgery be used to treat parkinsons

A

Deep brain stim (pacemaker like unit which can be tunred on or off) Tallatomy (removal of parts of the thalamus) Pallidotomy (destruction of parts of the globus pallidus)

239
Q

Adult SCs lack

A

Telomerase

240
Q

What happens is S phase

A

Synthesis of DNA

241
Q

What does the epiblast give rise to

242
Q

What cell type does the non cycling precurrsors give rise to

A

Cycling secretory progenitors

243
Q

At which end is FGF gdt greatest during somiteogenesis

A

Spinal chord end

244
Q

When is cyclin A found

245
Q

NOn motor symptoms

A

Orthostatic hypotension Respiratory problems Bowing of shoulders Swelling fo the feet Increased sweating

246
Q

Key difference between tissue specific and PSCs in vitro

A

Adult are difficult to isolate and maintain in vitro

247
Q

CELL SURFACE MARKERS MOUSE ES CELLS SSEA1 SSEA3/SSEA4 ALKALINE PHOSPHATASE

A

SSEA1 Y SSEA3/SSEA4 N ALKALINE PHOSPHATASE Y

248
Q

How does somiteogenesis occur

A

High FGF signalling at posterior end keeps cell in proliferating state FGF and RA give posterior identity and anti WNTs maintain anterior identity

249
Q

Effect B-catenin

A

Turns on target genes displaces co repressors TCF LEF

250
Q

STEM CELL GENES HUMAN ES CELLS OCT4 NANOG SOX2 KIF4 DPPA3 REX1 GBX2 FGF5

A

OCT4 Y NANOG Y SOX2 Y KIF4 Y DPPA3 Y REX1 Y GBX2 Y FGF5 N

251
Q

Steps for generation of a transgenic

A

Transfect ES cells Genotype targetted cells and select the correctly modified cells ES cells microinjected into the host blastocyst Blastocyst transferred into pseudo pregnant mouse Chimeras Cross for germline transmission for homozygosity

252
Q

Describe the intrinsic pathway of apoptosis

A

DNA damage + p53 Mt release cyt-c Initiator caspase 9 Effector caspase 3

253
Q

What happens in G1

A

Cell grows Ribosomes and RNA production

254
Q

What two gradients induce neuralation

A

SHH and BMP

255
Q

Three divisions of the human auditory system

A

Outer ear Middle ear Inner ear

256
Q

What are there potentially in the human ear

A

Stem cells

257
Q

Which electrical implant essentially acts as the hair cells

A

Cochlear implant

258
Q

Potential risks of using SCs in regenerative medicine

A

Lack of efficacy Side effects Cancer Rejection

259
Q

Properties of human artificial chromosome

A

Self replicating and sequenced Behaves as a stable chromosome Independent from chromosome of the host cell

260
Q

Cyclin B pairs

261
Q

Advantages of TAT fusion

A

Non integrating

262
Q

What is transdifferentiation

A

Trying to convert one cell type to another Can be done with the correct factors

263
Q

What happens at the G1 checkpoint

A

Is cell big enough and is the environment favourable

264
Q

Cyclin D pairs

BMS114 Stem Cells and Society (1 decks)

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