earticle

논문검색

Induced Pluripotent Stem Cell Generation using Nonviral Vector

초록

영어

Induced pluripotent stem (iPS) cells have been generated from mouse and human somatic cells by etopic expression of transcription factors. iPS cells are indistinguishable from ES cells in terms of morphology and stem cell marker expression. Moreover, mouse iPS cells give rise to chimeric mice that are competent for germline transmission. However, mice derived from iPS cells often develop tumors. Furthermore, the low efficiency of iPS cell generation is a big disadvantage for mechanistic studies. Nonviral plasmid‐based vectors are free of many of the drawbacks that constrain viral vectors. The histone deacetylase inhibitor valproic acid (VPA) has been shown to improve the efficiency of mouse and human iPS cell generation, and vitamin C (Vc) accelerates gene expression changes and establishment of the fully reprogrammed state. The MEK inhibitor PD0325901 (Stemgent) has been shown to increase the efficiency of the reprogramming of human primary fibroblasts into iPS cells. In this report, we described the generation of mouse iPS cells devoid of exogenous DNA by the simple transient transfection of a nonviral vector carrying 2A‐peptide‐linked reprogramming factors. We used VPA, Vc, and the MEK inhibitor PD0325901 to increase the reprogramming efficiency. The reprogrammed somatic cells expressed pluripotency markers and formed EBs.

목차

ABSTRACT
 INTRODUCTION
 EXPERIMENTAL PROCEDURES
  Mouse Embryonic Fibroblast Isolation and iPS Cell Culture
  Plasmid Preparation and Infection
  AP Staining
  Immunocytochemistry
  Quantitative Real-Time Polymerase Chain Reaction (Real-Time PCR) Analysis
  Embryoid Body Formation Analysis
  Statistical Analyses
 RESULTS
  Induced Pluripotent Stem Cell Generation using a Nonviral Vector
  Virus-Free Induced Pluripotent Stem Cells Express Pluripotency Marker Gene
  Characterization of iPS Cells Generated using Nonviral Vectors
  In Vitro Differentiation
 DISCUSSION
 REFERENCES

저자정보

  • Si Jun Park School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University
  • Mi Jung Shin School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University
  • Byoung Boo Seo Dept. of Animal Resources, College of Life & Environmental Science, Daegu University
  • Humdai Park Dept. of Biotechnology, College of Engineering, Daegu University
  • Du Hak Yoon School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University
  • Zae Young Ryoo School of Life Science and Biotechnology, College of Natural Sciences, Kyungpook National University

참고문헌

자료제공 : 네이버학술정보

    함께 이용한 논문

      ※ 기관로그인 시 무료 이용이 가능합니다.

      • 4,000원

      0개의 논문이 장바구니에 담겼습니다.