參訪 Atit

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參訪 Atit

  1. 1. ATIT- Animal Technology Institute Taiwan . ; 972004206; 2009/11/9 iPSC ppt
  2. 2. 1970 2001 induced pluripotent stem cells 2006 Shinya Yamanaka protein-induced Pluripotent Stem Cells 2009 Hongyan Zhou 1962 JB Gurdon Xenopus 2009 iPSC piPSC ”piPSC iPSC” Xenopus laevis 1962, JB Gurdon 2009, Zhou et al. egg 1962 John Gurdon nuclear transplant ” ” (egg) nuclear blastula, gastrula endoderm ” ” transplantation cloning 1950 tadpole (JB Gurdon, 1962) nuclear transplantation 2009 Albert- Lasker Basic Medical Research Award 1962 paper oct-4 2003 JB Gurdon (thymus) Xenopus oocytes X. laevis paper Xenopus oocytes oct-4 oocytes oct-4 knockout oct-4 transcription factor POU5F1 POU family 2006 Shinya Yamanaka Oct3/4, Sox2, c-Myc, Klf4 nuclear reprogramming( ) retroviral fibroblast hES medium colony iPSC Yamanaka Fbx15 iPSC 2
  3. 3. DNA eukaryotic cell 1. ES cell teratoma 2. DNA methylation Transfection: 3. chimera 1. electroporation 2. calcium phosphate, 4. factor c-Myc oncogene DEAE-dextran cell DNA 3.Lipofection 2007 Yamanaka iPS cell ES 4.Microinjection( 1962JB Gurdon ) cell Nanog selection marker Fbx15 DNA methylation chimera Transduction: retrovirus Hongyan Zhou recombinant protein piPSC reverse-transcribed iPSC DNA gene manipulation DNA Zhou fibroblast piPSC in vitro in vivo pluripotency poly-arginine protein CPP(cell transduction domain transcription factors C-terminus CPP penetrating peptides) cell lysine arginine amino acids cell paper gene transcribe translated 1962 JB Gurdon !"#$%&'()* +,-.(X.laevis)/0 12345 1998 JA Thomson 2003 MN„…e†|vz~x}‡z~ 1998 JA Thomson ESC JB Gurdon et al. ˆ‰[ 67&89:;<=> $%1?@AXenopus BCDoct-40E8(F 2007 Thomson Yamanaka GHIJ$KL ES 2006 Yamakana et al. Yamanaka 2009 review HIA]^123_` a&bcde(fdgh iPS teratoma ijk0lmnop/ 2007 UqrsYZ[ Yamakana et al. Cell Stem Cell `YZ[—˜(F”)* Zhou et al., piPSC c…0šY›‚x›|vz~•œ 2007 iPSC Yamakana et al. Š‹ŒG•aŽ•*…e$ 2009 %•Yw‘’}{(“G”•– t*uvwxy/z{|{}~Yxw• Yamakana et al. €v•{‚/UqƒsYZ[ 2007 •{žY{Ÿ ¡¢YZ[0£¤ JA Thomson ¢¥¦§•r¨©ªa XYZ[«I¬- 2009 `YZ[—˜(r™67 *…•/ Zhou et al. *MNO@PQR0S T/UVWXYZ[ 3
  4. 4. 2007 Yamakana iPSC 1. low reprogramming efficiency OSKM factors pluripotent cell 2006 paper 0.01–0.5% 2007 Yamakana 2006 Fbx15 1/10 0.001–0.03% factors Oct4, Sox2, Klf4, and 2. genomic alterations due to viral integration reverse transcribe OSKM DNA DNA c-Myc 3. c-Myc Klf-4 oncogene telomere iPSC/piPSC telomerase (~50 cycles) ES cell telomerase telomere 2007 Yamanaka Nanog selection marker iPSC hTERT(human Telomerase Reverse Transcriptase) hES(human ES)cell line hES iPSC 60 clone cell doubling time • 46.9 ± 12.4 (201B2) • 47.8 ± 6.6 (201B6) • 43.2 ± 11.5 (201B7) • hES doubling time clones iPSC telomere GFP(Green Fluorescent Protein) Chimera GFP transfection Yamanaka paper GFP ” ” GFP Lasker Nobel Prize 4
  5. 5. 2007 stem cell gene targeting in mice Bibliography 1. Campbell NA, Reece JB. BIOLOGY. 6th ed. Benjamin iPSC piPSC Cummings; 2001. oncogene Literature letter PNAS 1. , , , (2008) pp ”Distinguishing Cancerous iPSCs from ESCs” 20-26 iPSC ESC 2. Zhou et al., Generation of Induced Pluripotent Stem Cells iPSC Using Recombinant Proteins, Cell Stem Cell (2009) pp. 1-4 3. JB Gurdon, The Developmental Capacity of Nuclei taken from Intestinal Epithelium Cells of Feeding Tadpoles, J. Cancerous iPSC Embryo. exp. Morph., (1962) pp. 622-40 4. JB Gurdon et al., Nuclei of Adult Mammalian Somatic Cells Are Directly Reprogrammed to oct-4 Stem Cell Gene Expression by Amphibian Oocytes, Current Biology (2003) pp. 1206–1213 5. S Yamanaka et al., Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors, Cell, (2006) pp. 663-676 6. S Yamanaka et al., Generation of germline-competent induced pluripotent stem cells, Nature, (2007) pp. ... 313-317 20 60~80 7. JA Thomson, Embryonic Stem Cell Lines Derived from Human Blastocysts, Science, (1980) pp. 1145-47 8. S Yamanaka et al., Embryonic Stem Cell Lines Derived from Human Blastocysts, Nature Biotechnology, (2009) pp. 1-3 9. S Yamanaka et al., Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors, Cell, (2007) pp. 861-872 10. JA Thomson et al., Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells, Science Express, (2007) pp. 1-4 11. RM Marion et al., Telomeres Acquire Embryonic Stem Cell Characteristics in Induced Pluripotent Stem Cells, Cell Stem Cell, (2009) pp. 141-154 or 5

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