Induced Pluripotent Stem Cell Generation using Nonviral Vector

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 35권 / 4호
ㆍ저자명 : Si Jun Park, Mi Jung Shin, Byoung Boo Seo, Humdai Park, Du Hak Yoon, Zae Young Ryoo

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

영어 초록

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.

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