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Advanced molecules, membranes and cells > Genetic Manipulation 1 > Flashcards

Genetic Manipulation 1 Flashcards

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Organic Chemistry 101 flashcards
1
Q

Pluripotency

A

The ability of a cell to contribute to any tissue in the body

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2
Q

pluripotent teratoma examples

A

sacrococcygeal teratomas in males 1 in 35 000

ovarian teratoma

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3
Q

what are teratomas?

A

tumours formed by germ cells that form oocytes and sperm

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4
Q

ovarian teratomas

A

usually benign and contain differentiated tissues

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5
Q

testicular teratocarcinoma

A

EC cells which are pluripotent

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6
Q

Embryonic stem cells

A

Pluripotent stem cells derived from inner cell mass of blastocyst of pre-implantation embryos

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7
Q

how many cells in the pre-implanted blastocyst?

A

50-150

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8
Q

Who discovered ES cells?

A

martin evans and matthew kaufman

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9
Q

What can you find out about ES cells in culture?

A

check expression of ICM/stem cell genes oct 4 and nanog for pluripotency
cell surface markers eg SSEA1
check sex and karyotype - SRY and HPRT

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10
Q

What is LIF used for?

A

leukaemia inhibiting factor prevents ES cell differentiation

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11
Q

What happens when ES cells are introduced into an adult mouse?

A

form teratomas

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12
Q

what happens when ES cells are re-introduced into a blastocyst?

A

contribute to all parts of embryo and form a chimeric mouse

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13
Q

oct 4

A

transcription factor expressed in ICM cells

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14
Q

what happens if there are no oct 4 genes?

A

develop into blastocyst but then die

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15
Q

what happens if there are no nanog genes?

A

lose pluripotency and develops extra embryonic tissue - parietal endoderm

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16
Q

What happens if LIF is withdrawn from ES cells grown on dishes or in suspension?

A

form ball like embryoid bodies

17
Q

embryoid bodies

A

contain differentiating cells of 3 embryonic lineages and are trying to form an embryo

18
Q

what may embryoid bodies also contain?

A

cardiomyocytes that can beat

19
Q

How can we get fully GM mice from ES cells?

A

genetically manipulate cells - mutation/new genes
reintroduce into mice - inject into ICM
chimeric mice - breed these together

20
Q

How is DNA introduced into cells?

A

bathe cells in media containing the DNA and pass shock through them - electroporation

21
Q

Electroporation

A

introduced DNA into cells by making small holes in the cell membrane

22
Q

What is done to the female mouse before implanting the ES cells?

A

mated with vasectomised mouse to allow uterus to swell and vascularise?

23
Q

How to tell the difference between host blastocyst and donor ES cells

A

different strains of mice and different coat colours

Advanced molecules, membranes and cells (20 decks)

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