All Lectures Flashcards
Lecture 1-18
1
Q
Stage 3 trials
A
Prolonged
2
Q
5 vector based approaches
A
Transient transfection Adenovirus Floxed vectors Transposon mRNA
3
Q
Which methods may be used to identify iPSCs
A
Epigenetics Protein and gene expression Differentiation Morphology
4
Q
What cell types does the myeloid progenitor give rise to
A
Neutrophils Basophils Eosinophils Monocytes Erythrocytes
5
Q
Name some of the functional tests for PSCs
A
Teratoma form Embroid bodies Single cell replating Spin EB and growth factors
6
Q
Stage 1 trials
A
Safety
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
8
Q
DEVELOPMENTAL POTENTIAL MOUSE ES CELLS TERATOMA CHIMERA
A
TERATOMA Y CHIMERA Y
9
Q
In the mouse embryo what is the fate of the inner cells
A
Form the epiblast then the embryo
10
Q
What is IRES used for
A
Knock in Sequence when in transcript ribosome can bind so two different proteins are produced
11
Q
As the primitivie streak extends what becomes more anterior
A
Node
12
Q
What is the point of no return in the cell cycle
A
Restriction point
13
Q
When is cyclin E found
A
G1/S
14
Q
Chemical bases approaches
A
VPA 5-AzaC
15
Q
Describe gastrulation
A
Epiblast cells migrate through the primitve streak emerging and endoderm and mesoderm this produces the 3 germ layers
16
Q
Advantages of floxed based delivery
A
Efficient Can be removed
17
Q
What are tissue specific SCs also known as
A
Adult
18
Q
As the embryo grows what are pushed more anteriorly
A
AVE AND DVE
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
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
21
Q
What can be said about the colonogenic capacity of hES cells
A
Low
22
Q
How would you specify DA neurons
A
Supress mesoderm and promote ectoderm Inhibit BMP and TGFB
23
Q
Describe the cell cycle of PSCs
A
G1 shorter Self renewal maintained in G2/S
24
Q
How are embroid bodies used
A
Induce differentiation and examine the 3D structure
25
Mutations in BMP TGF-B can cause
Cancer
26
What virus can transduce both dividing and non dividing
Can transduce both dividing and non-dividing cells
27
Disadvantages of adenovirus based delivery
Repeated infection required for certain cell types Delayed kinetics of reprogramming
28
How can you tell RPE cells in suspension
Pigmented
29
Implications of the ASC model
Cells flow through in unidirectional manner Differentiated cells become restricted Ordered and irreversible decisions
30
Descirbe the restriction point
CDK4/Cyclin D --\> Phsophrylation of E2F and pRb CDK2/Cyclin E --\> diphosphorylation of E2F and pRb Dissociation E2f enters the nucelus
31
Stages for drug disocvery
Target identification Lead selection Lead optimise Pre clinical development Clinical trials phase 1 2 3
32
Give of an example of a stem cell with a medium turnover
Red blood cells Cardiomyocytes
33
Describe physiological assay
Low throughput difficult assay physiological
34
Which AMD variant is more common
Wet 90%
35
RESPONSE TO FACTORS MOUSE EPISC LIF NODAL/ACTIVIN FGF2
LIF N NODAL/ACTIVIN Y FGF2 Y
36
What happens at the metaphase-anaphase transition
Check whether all chromosomes attached to the spindle
37
Where in each signalling pathway may feedback act
Every step
38
Which two progenitors are produced by the mPSCs in the blood
Lymphoid progenitor and the myeloid progenitor
39
Stage 2 trails
Efficacy
40
INK4 family
Inhibit CDK4
41
CELL SURFACE MARKERS MOUSE EPISC SSEA1 SSEA3/SSEA4 ALKALINE PHOSPHATASE
SSEA1 Y SSEA3/SSEA4 N ALKALINE PHOSPHATASE N
42
Symptoms of AMD
Trouble disconcerting colours Blurred vision Decrease in visual acuity Slow recovery after exposure to bright lights
43
What is found between the RPE cells and the choroid
Bruchs membrane
44
Steps for induction of pluripotency
Somatic cells differentiated Viral transduction Colony formation IPS cells
45
Where does TFIID bind
The TATA box
46
Descibe activation of an RTP
Autophosphorylate tyrosine residues
47
Summarise ES cells
From embryo High prolif Highly potent High availability
48
Which types of viral transduction could be used to treat AMD
OCT4 SOX2 KLF4 C-MYC LIN28 NANOG
49
How would you specify ectoderm
Block TGF-B and BMP signalling
50
What method of gene editting may be used to bolt on other proteins
CAS9 defficient nuclease
51
Describe a normally functioning basal ganglia
DA produced by cells in the SN which affect other areas of the dorsal striatum
52
How can cultured PRE cells from post mortem material be used to treat AMD
In vitro human RPE retain sight with transplant into rat Can be tested with rodent head tracking
53
How can we stimulate sensory regeneration in the human ear
Need to remove inhibition of the stem cells
54
What is a TE
Transposable element which can change place in the genome
55
KIP family
CDK inhibitor protein
56
Describe the JAK STAT pathway
Cytokine receptors activate the JAK tyrosine kinase, phosphorylate themselves and other receptors STAT gene reg proteins Activated STATS dimerise and migrate to the nucleus
57
Give examples where human cell der cells lines are useful, which situations may they not be useful in
Useful in cancer BUT not if you wanted to study the very early stages of tumourigenesis
58
Functions of normal RPE cells
Regeneration of bleaches opsins Transepithelial transport Absorption of light Protection from photooxidation Secretion of growth factors Phagocytosis
59
3 germ layers
Endo/Ecto/Mesoderm
60
What virus can only tranduce dividing cells
Retrovirus
61
How can mRNA used to deliver
Exogenous RNA causes an ancient antiviral response causing degredation Need drugs to supress this and 5' protected mRNA
62
By what movement do the cells move through the primitve streak
Involution
63
How may cell replacement be used to treat parkinsons
Human foetal transplant (cells in the human foetus which make and release dopamine) Cells that make DA (can be similar or not (RPE cells)
64
What can be said about RA gradients during somitogenesis
Highest in the middle tails down either side
65
What is the effect of TGF-B and BMP signalling
They exert proliferative signals to cells
66
How are proteins degraded
Tagged with ubiquitin Enters proteasome and degraded
67
DEVELOPMENTAL POTENTIAL HUMAN ES CELLS TERATOMA CHIMERA
TERATOMA Y CHIMERA N/D
68
What project radially
Lysine rich N terminus tails of core histones
69
Broad classes of ways in which hearing loss can be prevented
Anti oxidants Anti apoptopic drugs NMDA blockers Growth factors
70
How can teratoma form be a test for PSCs
Should form all 3 of the germ layers
71
What is the frequency range of human speech
2-7 kHz
72
Advantages of transposon based delivery
Efficient Mark ffree excision
73
3 fundamental properties of stem cells
Potency Self renewal Differentiation
74
Side effects of DA agonists
Similiar to LDOPA side effects, can also cause hallucinations
75
What do the absoptive progenitors give rise to
Enterocytes
76
Describe the pathway of ASCs in the muscles
Satellite cell Myoblast Myocyte Myotube
77
Advantages of mRNA based delivery
No viral componenets No genomic integration
78
What determines the cell type
The repitoire of genes switched on
79
Advantages to virus based delivery
Efficient delivery Should be silenced
80
What is special about cyclin D
Direct link between the EC environment and the cell cycle Target of many growth factors
81
4 signalling centres in embryos
Post epiblast Anterior visceral endoderm Exterior embyonic endoderm Node
82
Issues with Talens/ZFN
Protein:DNA interactions Hard to design
83
Disadvantages of floxed based delivery
Genomic integration: risk of insertional mutagenesis Expresion often maintained in iPSCs Leaves a mark
84
What happens when hair cells are damaged in chicks
Transdifferentiation of supporting cells Proliferative regeneration
85
What happens in G2 phase
Cell checks fidelity of the DNA and preparation for nuclear division
86
Disadvantages of virus based delivery
Genomic integration: risk of insertional mutagenesis Expression often maintained in iPSCs: increased risk of tumours
87
CELL SURFACE MARKERS HUMAN ES CELLS SSEA1 SSEA3/SSEA4 ALKALINE PHOSPHATASE
SSEA1 N SSEA3/SSEA4 Y ALKALINE PHOSPHATASE Y
88
In ectodermal patterning what determines the AP axis What drives neural differentiation
FGFs BMP
89
What is 2A used for
Cleaves protein and is inserted
90
What is special about the +4 LRCs
Can revert back to the LGR 5+
91
What do the myo-SC give rise to
Myoepithelium
92
What can be seen in parkinsons patients
Lewy bodies made of alpha synuclein
93
Define neuralation
FOlding of the neural plate to give the nerural tube
94
Why is atoh1 not expressed in the supporting cells
Intracellular domain of notch is cleaved off in the supporting cells and enters the nucelus allowing the Hes to repress Atoh1
95
Route to specifying midbrain DA neuroens
BMP inhib WNT SHH and Low WNT
96
Describe the fovea
Closely packed cones Accurate vision
97
Stages for IVF/Embryo transfer
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
98
Which medications may be used to treat parkinsons
Dopemaine agonists MAO/COMT inhibitors L-DOPA
99
General ASC model
Stem cell gives rise to progenitors Progenitors expand Hardwired hierarchy Asymm division ASC divide few times to protect themselves from muations
100
Purposes of using drugs to treat disease
Reverse disease Prevent
101
Causes of hearing loss
Noise Genetics Ageing disease
102
What does CRSIPR use to target correct DNA sequence
Uses guide RNA
103
EXCEPTION TO THE MODEL Flow isnt always unidirectional
Loss of basal stem cells Clara cells de differentiate to replace
104
What cell types do the lymphoid progenitor give rise to
NK cells B and T lymphocytes
105
Give of an example of a stem cell with a lifetime turnover
Lens Oocytes
106
Descirbe the niche triangle
Cell:soluble factors Cell:ECM environment Cell:Cell
107
STEM CELLS GENES MOUSE ES CELLS OCT4 NANOG SOX2 KIF4 DPPA3 REX1 GBX2 FGF5
OCT4 Y NANOG Y SOX2 Y KIF4 Y DPPA3 Y REX1 Y GBX2 Y FGF5 NO
108
What SCs would come from the blastocyst
mESC hESC EpiSC Naive human SC
109
Describe the macula
Functional centre of the retina responsible for 20/20 vision
110
At which 3 places does histone acetylation occur
Aetylation of lysines Phosphorylation of serine 10 in H3 Mono/di/tri methylation of lysine and arginine in core histone N terminus tails
111
Describe the process for deriving the ES cells
Hatched blastocyst Antitrophectoderm and complement Leaves innner cell mass
112
What do stem cell properties allow
Regeneration Development - building embryos and tissue
113
What do the Lgr 5+ CBC give rise to
Non cycling precurrsors +4 LRC and absorptive prgenitor
114
Motor symtpoms
Slowness of movement Decreased ability to perform spontaneous movements Postural changes Shuffling gait Resting tremor
115
Which viruses may be used to induce pluripotency
OCT4 SOX2 KLF4 C-MYC LIN28 NANOG
116
When active what does TGF act
SMAD 2 and 3
117
Malfunction of apoptosis can lead to
Neurodgen disease Ischameic stroke Autoimmune disease HIV Cancer
118
Describe the niche in muscles
Between muscle fibres and myofibres
119
What tissues from the endoderm
Pancreas Liver
120
What were the two 'senses' for the origin of the stem cell
It is the unicellular origin of life The fertellised oocyte is the origin of all cells
121
When active what does BMP act
SMAD 1 5 8
122
What else do transcription activators bind to
The mediator complex
123
What can be said about PSCs when growing in vitro
Will proliferate indefinetly
124
Describre high throughput screening
High througput Simple Not single cell so would need a target or reported
125
RESPONSE TO FACTORS HUMAN ES CELLS LIF NODAL/ACTIVIN FGF2
LIF N NODAL/ACTIVIN Y FGF2 Y
126
What does a primed cell mean
Expresses lineage markers
127
What does the haematopoetic stem cell give rise to
Multipotent stem cells
128
Properties of ES cells
Indef prolif pot Carry particular markers Pluripotent Not transformed but high telomerase Derv from inner cell mass of blastocyst Clonergenic Stable diploid karyotype
129
Why would iPSCs be used
Capture a diseased genotype OF A HUMAN POP OR DISEASED GENOYPE
130
Cyclin E pairs
CDK 2
131
What type of signalling is used between hair cells and the supporting cells
Notch
132
Give situations where cells may need to die by apoptosis
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
Effects of alternate splicing
One gene can create multiple proteins
134
How does RNAi work
Physically blocks translation
135
Positives of using PSCs to treat Parkinsons
Could be gorwn in large batches and immune matched
136
Summarise tissue specific stem cells
Low prolif From adult and juvenile and foetal organs Multipotent Hard to access and isolate
137
What occurs with dry AMD
Little no haemorhaging
138
What component parts of the BG
Nucelus accumbens Substantia nigra striatum Globus palidus Thalamus STN
139
Analysis methods of disease models
Antibody staining Cell counts Reporters
140
Descirbe the basal ganglia in Parkinsons patients
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
Disadvantages of transposon based delivery
Clones need to be checked for excision and rearrangements
142
Overall effect of acetylation of histones
Opens up the chromatin
143
STEM CELL GENES MOUSE EPISC OCT4 NANOG SOX2 KIF4 DPPA3 REX1 GBX2 FGF5
OCT4 Y NANOG Y SOX2 Y KIF4 N DPPA3 N REX1 N GBX2 N FGF5 Y
144
Descirbe some basic niche concepts
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
Give of an example of a stem cell with a quick turnover
Most epithelia Colon crypt cells
146
Problems with cell transplantation to treat AMD
Neural retinal restored to reveal RPE cell layer and transplanted onto the macula
147
Describe WNT signalling in the ON state
WNT binds fizzled LRP5/6 binding complex activates dischevelled which prevents destruction of B-catenin enters nucleus
148
Components and reasons for components in a human artificial chromosome
Telomere - maint integrity Rep origin - so can be replicated Centromere - segregation of chromosomes
149
Describe the extrinsic pathway of apoptosis
Death ligands Death receptors Initiator caspase 8 Effector caspase 3
150
What gene is expressed in the hair cells
Atoh1
151
What part of the brain is effected in Parkinsons patients
Basal ganglia
152
In the epiblast what does BMP induce
Induces lefty1
153
Describe the action of nitric oxide
Chem ubstable Small and uncharged Cytoplasmic receptor
154
Mouse EpiSC
From pregastrualtion emb Primed
155
Disadvantages of mRNA based delivery
Multiple rounds of transfection Lower levels of expression than when integrated
156
Two categories of parkinsons symtpoms
Motor dysfunction and non motor symptoms
157
Define a niche
Defined anatomical location promoting self renewal
158
Problems with cell transplantation to treat AMD
Restoration of central vision at the expense of area of peripheral vision Complicated operation Follow up surgery to remove compression medium
159
In chick what is present during gastrulation
Hensons node
160
What do the secretory progenitors give rise to
Goblet cell Enteroendocrine cell Paneth cell
161
What tissues from the mesoderm
Heart Vascular Blood
162
Describe the sets for creating a disease model
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
RESPONSE TO FACTORS MOUSE ES CELLS LIF NODAL/ACTIVIN FGF2
LIF Y NODAL/ACTIVIN N FGF2 N
164
How do dopamine agaonists work
Not metabolised into DA but mimic the action
165
What does transposase catalyse
Movement of the transposable element to another part of the genome
166
General principle of eukaryotic transcription
Transcription factors bind to enhancers Allows RNA polymerase to bind to promoter region
167
Process for induction of pluripotency
Start with somatic cells that are diff Colony form iPSC cells signal to ES cells by markers
168
What activates TGF-B
Nodal Activin-A TGF-B
169
What are the choices for stem cells
Differentiation Death Self renewal
170
How can ASC assist in repair
Stop themsevles aging Prevent mutations Supply large numbers of differentiated cells
171
Two conditions of using PSCs to treat Parkinsons
Would need to ensure differentiaed to the correct cell type Check physiologically mature
172
Protein based approches
TAT-fusion
173
Describe the niche found in the Distal tip cell in the C.elegans
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
Summarise iPSC properties
Pluripotent High prolif capacity Easily available From somatic cells
175
Disadvantages of VPA 5-AzaC
Very inefficient
176
Disadvantages of TAT fusion
Inefficient
177
What do the Lum SC cells give rise to, finish the pathway
Alveolar progenitor --\> Alveolar cell Ductal progenitor --\> Ductal luminal cell
178
TGF and BMP compete for
SMAD 4
179
Where is WNT AND TGF-B signalling restricted to
Post epiblast
180
What can be said about the efficiency of cloning
Decreases with each stage and in more complex organisms
181
Which enzyme is involved in siRNA
DICER
182
What is used to fuse the myotubes
Horse serum
183
Describe the valley model
Cellular states like rolling a ball down a hill Valleys act as barriers
184
Describe high contrast screening
Single cell Mid throughput Diffiucult assay
185
What happens at the G2 checkpoint
Is all DNA replicated Is the cell big enough Is environment favourable
186
What forms at the highest concentration of NODAL
The node
187
Two sources of PSCS
Reprogrammed From the early mammalian embryo
188
How can a cell transplant to treat AMD work
Autologous RPE transplant maitains/restores sight in humans affected
189
Problems of L-dopa treatment
Prolonged can caused dyskineasias
190
What tissues from the ectoderm
Non neural Neural
191
What happens if you remove a SC from the niche
Differentiation
192
Three types of elecrtical implants used what does each of them bypass
Auditory nerve inplant - bypasses the cochlear Brainstem implant - bypasses cochlear and auditory nerve Cochlear implant - bypasses the hair cells
193
In the mouse embryo what is the fate of the outer cells
Extraembryonic (placenta)
194
Describe WNT signalling in the OFF state
B-catenin associated with E-cadherin Targetted phosphorylation --\> ubiquitination --\> degredation of B-catenin
195
What happens in G0
Cell withdraw from the cell cycle, replication machinery is dismantled and cell enters a quiescent state
196
When is cyclin B found
G2/M
197
Cyclin A pairs
CDK 2/1
198
What is drunsen
ECM build up between Bruchs membrane and RPE cells of the eyes
199
What is the solution for AMD treatment
RPE cells in solution at high density + bare patch can be used to create an RPE cell coated patch
200
What did Weismann propose in 1892
Germ plasm continuity
201
MASC can give rise to (mammary gland)
Myo-SC Lum-SC
202
Describe PI3 kinase activation
Leads to activation of Akt
203
Problems with cultured RPE cells being transplanted from post mortem material to treat AMD
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
Why may CRISPR be preferred
RNA:DNA interactions Easier to design
205
Functions of dopamine
Related to mood Stimulates smooth and balance muscle movement
206
Steps for cloning
Take an adult somatic cell and remove nucleus Insert nucleus into an unfertilized oocyte Electric shock to stimulate uptake of the nucleus
207
Possible concepts of using stem cells to treat hearing loss
Can we differentiate ES cells to hair cells Transplanting Awakaken endogenous stem cells Do any stem cells exist in the inner ear
208
When is cyclin D found
G1
209
Advantages of VPA 5-AzaC
Very easy to use Cheap
210
How would you specify primitive streak cells
TFG-B and WNT
211
Describe the notch signalling pathway
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
More Shh in the brain gives a
More ventral cell fate
213
During cloning what happens
SOMATIC GENES OFF EMBRYO GENES ON DNA methylation back to the totipotent configuration Cell nucleus manipulated to reset the state
214
Advantages of adenovirus based delivery
Lower frequency of genomic integration
215
Advantages of using human PSCs
Differentiation to any cell fate Grow indefinetly Normal primary line
216
HES
Primed
217
How does L-dopa work
DA precurrsor which is able to cross the blood brain barrier and be metabolised to DA
218
EXCEPTION TO THE MODEL Not all SCs are multipotent
Epidermal basal cells ONLY make karatinocytes
219
Steroid hormones
Structurally similar to cholesterol Hydrophobic IC receptor
220
Hearing loss prevalence 17-30 31-40 41-50 51-60 61-70 71-80
1.8 2.8 8.2 18.9 36.8 60.2
221
EXCEPTION TO THE MODEL Not all cells are quiescent
E.g. in intestinal crypt cells SC are continuouskly dividing
222
Why is age a problem with AMD tratement
Aged bruchs membrane, RPE cells may fail to attach
223
At what stage are apical and basolateral domains established
Early morula
224
Mouse ES
From blastocyst Naive
225
What activates BMP
GDF3 Noggin
226
Characteristics of cancer
Loss of restriction point Normal cell cycle regulatory machinery lost Loss of contact inhibition Failure to respond to growth factors Cells cant arrest
227
Problems with using mouse cells lines
Physical dif Physiological dif Genetic dif SOme mutations in the BLM genes are fatal in mice but cause disease in humans
228
Characteristics of necrosis
Total cell lysis No vesicle formation Loss of integrity Swelling of the cytoplasm Disintegration of organelles
229
What occurs with wet AMD
Growth of new blood vessels within the retinal tissue and subsequent haemorrhage
230
Effects of DNA methylation
Directly blocks TFIID binding Recruitment of histone de-acetylases
231
What can Ras activate, describe this activation
MAP kinases M kkk M kk M k Gene expression/regulatory activity
232
What is the ASC in the small intestine crypt villus unit
Lgr 5+ CBC
233
How could PSCs be characterised by genetics
Imprininting X inactivation Methylation
234
Prevalence in 40+ 65+ 80+
0.4% 1% 10%
235
DEVELOPMENTAL POTENTIAL MOUSE EPISC TERATOMA CHIMERA
TERATOMA Y CHIMERA N
236
What are the 3 main types of non-integrating delivery of reprogramming factors
Vector based Protein based Chemical bases
237
Stratergies of generating disease models
Healthy PSCs then gene edit Grow indefinetly Somatic cells --\>\> iPSCs and then gene edit
238
How may surgery be used to treat parkinsons
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
Adult SCs lack
Telomerase
240
What happens is S phase
Synthesis of DNA
241
What does the epiblast give rise to
Ectoderm
242
What cell type does the non cycling precurrsors give rise to
Cycling secretory progenitors
243
At which end is FGF gdt greatest during somiteogenesis
Spinal chord end
244
When is cyclin A found
S and G2
245
NOn motor symptoms
Orthostatic hypotension Respiratory problems Bowing of shoulders Swelling fo the feet Increased sweating
246
Key difference between tissue specific and PSCs in vitro
Adult are difficult to isolate and maintain in vitro
247
CELL SURFACE MARKERS MOUSE ES CELLS SSEA1 SSEA3/SSEA4 ALKALINE PHOSPHATASE
SSEA1 Y SSEA3/SSEA4 N ALKALINE PHOSPHATASE Y
248
How does somiteogenesis occur
High FGF signalling at posterior end keeps cell in proliferating state FGF and RA give posterior identity and anti WNTs maintain anterior identity
249
Effect B-catenin
Turns on target genes displaces co repressors TCF LEF
250
STEM CELL GENES HUMAN ES CELLS OCT4 NANOG SOX2 KIF4 DPPA3 REX1 GBX2 FGF5
OCT4 Y NANOG Y SOX2 Y KIF4 Y DPPA3 Y REX1 Y GBX2 Y FGF5 N
251
Steps for generation of a transgenic
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
Describe the intrinsic pathway of apoptosis
DNA damage + p53 Mt release cyt-c Initiator caspase 9 Effector caspase 3
253
What happens in G1
Cell grows Ribosomes and RNA production
254
What two gradients induce neuralation
SHH and BMP
255
Three divisions of the human auditory system
Outer ear Middle ear Inner ear
256
What are there potentially in the human ear
Stem cells
257
Which electrical implant essentially acts as the hair cells
Cochlear implant
258
Potential risks of using SCs in regenerative medicine
Lack of efficacy Side effects Cancer Rejection
259
Properties of human artificial chromosome
Self replicating and sequenced Behaves as a stable chromosome Independent from chromosome of the host cell
260
Cyclin B pairs
CDK 1
261
Advantages of TAT fusion
Non integrating
262
What is transdifferentiation
Trying to convert one cell type to another Can be done with the correct factors
263
What happens at the G1 checkpoint
Is cell big enough and is the environment favourable
264
Cyclin D pairs
CDK 4/6
