研究人員發(fā)明一種新工具,，它在促成多能干細胞中基因的選擇性表達時,，不會影響已經(jīng)分化的細胞，這一新成果發(fā)表在日前在線出版的《自然—方法學》期刊上。
誘導多能干細胞（iPS）是通過程序化重組成熟細胞而制成的,，它們有分化成各種成熟細胞類型的潛能,，但程序化重組過程的效率非常低。
James Ellis和同事合作,，講述了一種可促成基因在iPS細胞中獨自表達的病毒載體如何提高了iPS細胞的選擇性,。通過一種抗生素耐藥性基因的表達，它們用這些運輸器將抗生素耐藥性告知給新出現(xiàn)的iPS細胞,。Ellis和同事從雷特氏綜合征患者和模型小鼠體內(nèi)取出人類和小鼠的iPS細胞,。雷特氏綜合征是一種神經(jīng)失調(diào)疾病。新方法將被證明可應(yīng)用于實驗室里的疾病研究,，因為它能簡單地分離出小鼠和人類的iPS細胞系,。
新方法很可能應(yīng)用于制作疾病的試管模型，而iPS細胞則是通過重組取自特定疾病患者的細胞衍生,。（生物谷Bioon.com）
生物谷推薦原始出處：
Nature Methods 6, 370 - 376 (2009) 26 April 2009 | doi:10.1038/nmeth.1325
Isolation of human iPS cells using EOS lentiviral vectors to select for pluripotency
Akitsu Hotta1,2, Aaron Y L Cheung1,3, Natalie Farra1,3, Kausalia Vijayaragavan4,6, Cheryle A Séguin1,2,6, Jonathan S Draper1,6, Peter Pasceri1, Irina A Maksakova5, Dixie L Mager5, Janet Rossant1,2,3, Mickie Bhatia4 & James Ellis1,2,3
Induced pluripotent stem (iPS) cells may be of use in regenerative medicine. However, the low efficiency of reprogramming is a major impediment to the generation of patient-specific iPS cell lines. Here we report the first selection system for the isolation of human iPS cells. We developed the EOS (Early Transposon promoter and Oct-4 (Pou5f1) and Sox2 enhancers) lentiviral vector to specifically express in mouse and human embryonic stem cells but not in primary fibroblasts. The bicistronic EOS vector marked emerging mouse and human iPS cell colonies with EGFP, and we used puromycin selection to aid the isolation of iPS cell lines that expressed endogenous pluripotency markers. These lines differentiated into cell types from all three germ layers. Reporter expression was extinguished upon differentiation and therefore monitored the residual pluripotent cells that form teratomas. Finally, we used EOS selection to establish Rett syndrome–specific mouse and human iPS cell lines with known mutations in MECP2.
1 Developmental and Stem Cell Biology Program, Toronto, Ontario, Canada.
2 Ontario Human iPS Cell Facility, SickKids, Toronto, Ontario, Canada.
3 Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
4 McMaster Stem Cell and Cancer Research Institute, Michael G. DeGroote School of Medicine, and Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
5 Terry Fox Laboratory, British Columbia Cancer Agency, and Department of Medical Genetics, University of British Columbia, Vancouver, British Columbia, Canada.
6 Present addresses: Department of Regenerative Cardiology, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid, Spain (K.V.), 7 Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, Ontario, Canada (C.A.S.) and McMaster Stem Cell and Cancer Research Institute, Michael G. DeGroote Centre for Learning and Discovery, Hamilton, Ontario, Canada (J.S.D.).
