成果報告書詳細
管理番号20120000000639
タイトル*平成23年度中間年報 ヒト幹細胞産業応用促進基盤技術開発/ヒト幹細胞実用化に向けた評価基盤技術の開発 ヒトES細胞の品質評価指標の開発 (住友ベークライト株式会社)
公開日2012/9/11
報告書年度2011 - 2011
委託先名住友ベークライト株式会社
プロジェクト番号P10027
部署名バイオテクノロジー・医療技術部
和文要約和文要約等以下本編抜粋:1. 研究開発の内容及び成果等
(1) 研究開発の必要性
多能性を有する幹細胞は様々な細胞に分化する能力を有している。適切に誘導を行うことで神経、心筋、膵臓β細胞など様々な細胞を得ることができる。このため、創薬における薬効評価や安全性薬理試験などの創薬スクリーニング、発生・分化や疾患メカニズムの解明、再生医療への応用など生命科学や医療への貢献が大きく期待されている。
英文要約METI/NEDO New Project: Prof. Norio Nakatsuji, Sub-Project Leader
Title: Beyond the Fusion of Academia and Industry in Japan: an Integrated System for High Quality and Large Scale Production of hPSCs and Derivative Cells
Introduction:
The aim of this NEDO (New Energy and Industrial Technology Development Organization) Project is the “Development of Core Technologies for Industrial Applications of Human Stem Cells”. In recent years, great progress has been made worldwide in developing drug discovery screening and cell therapy applications for hPSCs ( human pluripotent stem cells, such as hES cells). However, large scale PSC culture methods vary widely across the globe, and are in need of standardization. Initiated last year in Japan, this six-year national project supported by METI (the Ministry of Economy, Trade and Industry) aims to develop not only large scale production methods, but also the technologies necessary for stem cell evaluation. Eight companies and five academic research groups are collaborating in this effort.
Research Plan and Targets (2011?2015)
1. Development of robust, mass culture and cryo-preservation technologies for hPSC lines
(1) Synthetic, defined culture medium with low molecular weight chemical compounds
(2) Culture substrate and materials for large scale cell culture systems
(3) Closed and minimal risk cell culture system and 3D mass culture technologies
(4) Automated cryo-preservation technology for hPSCs
2. Development of quality evaluation and control for hPSC lines
(1) Development of profiling/evaluation hardware and reagents for quality control of hPSCs
(2) Establishment of pluripotency evaluation technology by development of robust differentiation-inducing protocols
This year, Sumitomo Bakelite’s mission in this project is development of evaluation method of hPSC lines by glycan profiling. Glycan phenotype of hPSCs is known to be altered during differentiation stage. During differentiation, hPSC-associated glycan features will be replaced by differentiated cell-associated structures. Thus hPSCs differentiation stage can be determined by glycan profile, however, specific glycan markers for hPSCs quality have not established yet. Our aim is to find out the correlation between hPSCs quality and glycan profiles.
At first, reproducibility of the glycan profiling method was confirmed using some model cell lines by BlotGlycoR Glycan Purification and Labeling kit. Next, we performed N-glycan profiling using MALDI-TOF MS and LC-MS on hPSC lines (KhES-1 and KhS-3). Data demonstrated that hPSCs have characteristic N-glycomes which changes during hPSCs passage. Some N-glycans were expected to be promising ‘quality markers’ of hPSCs.
In the next year, we will establish a high-throughput glycan profiling system using automated separation equipment. Using the high-throughput system, we will establish a reliable quality marker by quantitative and structural analysis of hPSCs glycans. We are also planning O-glycan and glycolipid glycan analysis in the next stage.
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