Summary
Summary Trends in Stem Cell Biology (NWI-BM073)
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Written summary of all lectures Stem Cell Biology including paper discussions. Images added for extra clarity.
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, pluripotencysstteemm cows
cells that differentiate into other In mammals there two broad types of stem cells
Biological can ,
a re :
embryonic stem cells which isolated from the
types of cells and can divide to produce more of ,
a re inner
the same
type of stem cells (self renewal) cell mass of blastocyst s ,
and adult stem cells
In vivo loss of potency
2nd lineage decision :
ecto ,
endo and mesoderm
to -4 potent differentiation placenta and organism pluripotent
1
to -4potent : trophoblast and inner cell mass not most interesting ; can also form extraembryonic tissue
pluripotent : capable of differentiation into all three germlayers
multipotent :
progenitor cells (hennapoetic cell fate decisions are epi genomic based
run , potent : differentiate into one cell only because all cells contain the same DNA
Types of pluripotent cells
-
Embryonic stem cells (ESCs) : cells derived from embryo pluripotency is transient in the embryo ,
-
Epiblast stem cells (EP1 5CS) : cells derived from epiblast in culture phenomenon you fix them
-
Embryonot carcinoma cells €6s) : cells from terato carcinoma to stop differentiating
-
Embryonic Germ cells (EG Cs ) : cells derived from primordial germ cells
-
Induced pluripotent stem cell (PSG) : cell made from somatic cell
Test for pluripotency
-
Multilineage differentiation
differentiate into and mesoderm In human multi
→ ecto ,
endo
lineage differentiation in vivo not possible .
Extensive proliferation For absolute proof teratoma with differentiated cells of
germ layers
-
→ cells keep could be performed
growing in mice
Known marker /
genes proteins
-
→
when present it is a pluripotent cell
-
Produce germ line fhimensm)
inject stem cell with EGFP epiblast and from cell
→ in
grow organism single
-
Teratoma
if tumor with different cell
→ see
types is
being made
,Application of ES cells
-
model for
embryonic development Replacing cells is useful in
→
study gene regulation and characteristics -
stroke : loss of muscle cells
-
regenerative medicine -
duchenne muscle
:
degeneration
→
grow large quantities and differentiate into any tissue -
Parkinson loss of dopamine
:
generating cells
of KO
-
generation mouse
all cells to effect tissues
→
grow mouse with mutation in see on
Dangers
disease model
graft rejection
-
-
→ differentiate into desired specialisation -
graft versus host
tetracarcinoma
-
cytotoxicity tests -
Maintain pluripotency
cultured cells in dish are prone to differentiate
to inhibit differentiation vitro
ways
:
in
-
Feeders + serum
→ classic ,
mouse
embryonic fibroblast + cow serum
Life and batch dependent
serum cow serum
very undefined
-
+ is
→ Leahemea inhibitory factor
-
zi + Lf
→ 2 kinase inhibitors + 41 , very defined
Regulatory network
3 have autoregulated loop and activate themselves and eachother
→
upregulate pluripotency and self renewal
-
→ down
regulate developmental regulators L1F and Bmpu activate
pathways that directly talk
stem cells often have master regulators to
pluripotency network
→ bind promoters and enhancehers
→
regulate on
many
levels to sustain pluripotency
chromatin structure
chromatin modifiers a re important in development EUchromat.in Heterochromatin
→ cells need to change for differentiation H>Kume's H3K gmez
chromatin modifiers can assist
reprogramming Hzkzbmez Hskgmez
→ make it faster efficienter . or make it fat H3K 7g mes Hukzomes
HsKghaAc methylation
ES cells
-
globally decondensed (accessible)
enriched active histone marks (
green)
-
in
-
loosely bound chromatin proteins
Differentiated cells
-
condensed heterochromatin
decrease
adkennttcchhrroommaak-unn-stab.ly
-
silent histone marks in
acetylation B3W
bound chromatin proteins of bound to histone proteins
segments DNA . ,
that and
have both
repressing activating
epigenetic regulators in the same
region
, R
eprogwwaaddddxnnggttoonn.be#ggeenneeEtxccramming
marbles sample the
landscape is a metaphor for
and
how
at rest
gene regulation
at the lowest point
modulates development
the cell fate
the slope
groove on
,
come
,
.
Chromatin state corresponds with potency
→
pluripotent cells have flexible chromatin structure
→ differentiated cells have compact and specific chromatin state
somatic cell nuclear transfer Cell
reprogramming
nucleus from somatic cell The of mature specialised cells
I.
get process reverting ,
2. transplant in
oocyte without nucleus into induced pluripotent stem cells .
Erasure of
cell development
→
zn nucleus will develop like
embryo epigenetic marks during germ .
-
Technically challenging
-
Oocytes are
necessary
keepsakes ggeennee
A attached to regulatory sequence
gene
a
of of interest Fluorescence expressed
gene .
is
when
gene is
being expressed .
criteria for pluripotency
Differentiation capacity •
unlimited
growth
In UW0
morphology
- •
teratoma's under skin nude pluripotency markers
→ when ipscs
injected
•
mice
-
in vitro •
differentiation capacity ( in vivo and vitro)
→
embryo d bodies look for all layers
•
chimera /
germline transmission (not human ipscs)
, :
germ
IPS Cs
generation
four transcription factors can convert somatic cell into 1ps
Sequential event of
reprogramming
1. Initiation : add transcription factors
→ epithelial transition
mesenchymal - :
morphology change
fibroblast repressed and IPSC
genes induced
→
genes are
2. maturation Pioneer ffaaettorr
3. stabilization Transcription factor that can
directly bind to
→ I Pscs are stable : cells do not
stay in non - differentiate state condensed chromatin .
Recreate other factors
trans and histone modification enzymes and
→
genes are not necessary
anymore methylation
05k as pioneer
i. octy can bind to closed chromatin
and recruits factors to open heterochromatin
2. establishes pluripotency network
gene
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