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Triploid pluripotent stem cells. R eporter : XueBinghua Date : 2011.06.15. Human oocytes reprogram somatic cells to a pluripotent state. Scott Noggle & Dieter Egli Nature 1 August 2011. Background. Limit : Legal and social considerations Different : iPS and blastocyst-derived stem cells

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triploid pluripotent stem cells

Triploidpluripotent stem cells

Reporter:XueBinghua

Date:2011.06.15

human oocytes reprogram somatic cells to a pluripotent state

Human oocytes reprogram somatic cells to a pluripotent state

Scott Noggle & Dieter Egli

Nature

1 August 2011

background
Background
  • Limit:Legal and social considerations
  • Different:iPS and blastocyst-derived stem cells

(gene expression,DNA methylation and differentiation potential)

  • Compromise: Genomic integrity

(de novo mutations and copy number variations)

r esults
Results:

1、Development fails after somatic

genome replacement

mii oocyte donor cell
38/50

43/43 (12 without)

Skin cells (♂T1D and ♂WT )

GFP

CAGGS promoter:(H2B):GFP

MII oocyte & Donor cell
developmental potential development arrests
Normal cleavage :22/31(71%)

Arrests at a stage of 6–10 cells

Parthenotes:13/52 ( 25%)

IVF:16/21(76%)

Developmentalpotential & Development arrests
slide12
Results:

2、Development arrests with

transcriptional defects

slide13
6-12cells

*ZGA starts at the 4–8 cell stage28, or on day 3 of development*Upregulatedtranscripts incomparison to unfertilized oocytes 761 transcripts upregulated at zygotic genome activation 124 (16%) were upregulated after genome exchange 62 (8%) were upregulated after amanitin treatment 536(70%) were upregulated at parthenotes*1,406 genes expressat higher levels in the skin donor fibroblasts than in MIIoocytesGADD45G is involved in stress-induced cell cycle arrest

slide14
Results:

3、The oocyte genome rescues development

slide18
Results:

4、Derivation of pluripotent stem cells

g lobal gene expression analysis up and down
463/1327

670/1327

Global gene expression analysis &Up and down

*463:Lin28a,Oct4,Sox2, Nanog ,Lefty1

*670:FAP, PAPPA, MMP3,CTHRC1

SNAI2

*28 genes and 24 genes were expressed at higher levels in soPS2 and soPS1 than NYSCF1

neuropeptide galanin, SRY, rex1 ,

neuronatin, NODAL, Cerberus 1 ,

Lefty2

slide22
Results:

5、Epigenetic reprogramming

schematic of genome wide snp capture and allelotyping
Schematic of genome-wideSNP capture and allelotyping

Use a genome-wide digital allelotyping approach to distinguish gene expression from the somatic cell-derived genome and the oocyte-derived genome in soPS cells

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