論文 - 詳細
| RRC ID | 36642 |
|---|---|
| 著者 | Nishishita N, Shikamura M, Takenaka C, Takada N, Fusaki N, Kawamata S. |
| タイトル | Generation of virus-free induced pluripotent stem cell clones on a synthetic matrix via a single cell subcloning in the naïve state. |
| ジャーナル | PLoS One |
| Abstract |
CD34+ cord blood cells can be reprogrammed effectively on dishes coated with a synthetic RGD motif polymer (PronectinF®) using a temperature sensitive Sendai virus vector (SeV TS7) carrying reprogramming factors OCT3/4, SOX2, KLF4 and c-MYC. Dish-shaped human ES cell-like colonies emerged in serum-free primate ES cell medium (supplemented with bFGF) in 20% O2 culture conditions. The copy numbers of SeV TS7 vectors in the cytoplasm were drastically reduced by a temperature shift at 38°C for three days. Then, single cells from colonies were seeded on PronectinF®-coated 96-well plates and cultured under naïve culture conditions (N2B27-based medium supplemented with LIF, forskolin, a MAPK inhibitor, and a GSK inhibitor in 5% O2) for cloning purpose. Dome-shaped mouse ES cell-like colonies from single cells emerged on PronectinF®-coated dishes. These cells were collected and cultured again in primate ES cell medium supplemented with bFGF in 20% O2 and maintained on PronectinF®-coated dishes. Cells were assessed for reprogramming, including the absence of residual SeV and their potential for three germ layer differentiation. Generation of virus-free induced pluripotent stem cell (iPSC) clones from single cells under feeder-free conditions will solve some of the safety concerns related to use of xeno- or allogeneic-material in culture, and contribute to the characterization and the standardization of iPS cells intended for use in a clinical setting. |
| 巻・号 | 7(6) |
| ページ | e38389 |
| 公開日 | 2012-1-1 |
| DOI | 10.1371/journal.pone.0038389 |
| PII | PONE-D-12-04345 |
| PMID | 22719883 |
| PMC | PMC3374798 |
| MeSH | Animals Cells, Cultured Clone Cells / cytology* Clone Cells / immunology Culture Media Genetic Vectors Humans Karyotyping Kruppel-Like Factor 4 Mice Pluripotent Stem Cells / cytology* Pluripotent Stem Cells / immunology Primates Reverse Transcriptase Polymerase Chain Reaction Sendai virus / genetics |
| IF | 2.74 |
| 引用数 | 24 |
| WOS 分野 | MULTIDISCIPLINARY SCIENCES |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 14 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 研究用ヒト臍帯血幹細胞 | 研究用ヒト臍帯血幹細胞 |
| ヒト・動物細胞 | khES-1(HES0001) |