MCB 252, Part 2 Genetic inference, lecture 15 The 小型断路器2522的遗传推理、讲座15.pptVIP

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2017-03-08 发布于上海
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MCB 252, Part 2 Genetic inference, lecture 15 The 小型断路器2522的遗传推理、讲座15.ppt

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MCB 252, Part 2 Genetic inference, lecture 15 The 小型断路器2522的遗传推理、讲座15

Putting all 24 genes into MEFs “reprograms” Takahashi and Yamanaka (2006) Cell 126, 663-676 FBX15: an ESC-specific gene; only expressed in ESCs Viral promoter bgeo: G418 (an antibiotics that kills the cells) resistance gene So, cells can survive only when they become ESC-like cells Takahashi and Yamanaka (2006) Cell 126, 663-676 Narrowing down the candidates Oct4 (14) Sox2 (15) Klf4 (20) Myc (22) Takahashi and Yamanaka (2006) Cell 126, 663-676 iPS cells are pluripotent Pluripotency markers EB formation Teratoma formation - Saw the same thing with tail-tip fibroblasts Takahashi K., et al. (2007) “Induction of pluripotent stem cells from adult human fibroblasts by defined factors.” Cell 131, 861-72. OCT4, SOX2, KLF4, MYC Yu J., et al. (2007) “Induced pluripotent stem cell lines derived from human somatic cells” Science 318, 1917-1920. OCT4, SOX2, NANOG, LIN28 Park I.H., et al. (2007) “Reprogramming of human somatic cells to pluripotency with defined factors” Nature 451, 141-146. OCT4, SOX2, KLF4, MYC How about human cells? Adapted from: Gepstein. Circ Res 2002 /info/media/DSC_1187.jpg Translation Derivation Regenerative Medicine Stem Cell Biology Differentiation Tissue morphogenesis Cellular therapies Propagation Scale Up Quantitative, systematic approaches Quality control Stem cell-based therapy iPSCs Human somatic cells “Personalized medicine” Pitfalls with iPSCs Low efficiency of derivation 0.1% Use of C-myc Transgene integration Are they really the same as ESCs? - Are all four genes expressed in the same cells? Approach: Using a single retroviral or lentiviral vector instead of four vectors (2A peptide) Somers A, et al 2010, Stem Cells (STEMCCA Cre-Excisable lentivector) Staerk, J et al, 2010, Cell Stem Cell (T cells and myeloid cells) Chemical complementation (e.g., with small molecules such as VPA) to replace C-Myc Other compounds: Vitamin C, sodium butyrate, ALK5 inhibitor(*, mESC medium), Apigenin and Luteolin (E-cadherin enhancing)