成果報告書詳細
管理番号20130000000276
タイトル*平成24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」 (住友ベークライト株式会社)
公開日2014/8/20
報告書年度2012 - 2012
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
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
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 has been confirmed using some model cell lines by BlotGlyco® 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.
This year, we analyzed N-glycans on hPSC lines (KhES-1 and KhS-3) using LC-MS. The major glycans on the cell lines were high mannose type glycans. These findings are in accordance with previous studies in hPSC line (H1). Concerning the glycan analysis of the discrimination of the early and late passages of the PSCs, high mannose type glycans leaned to contribute the discrimination. We have found some of the N-glycans candidates that may be of markers to determine the (early / late) the difference between passage of undifferentiated ES cells using LC-MS. Moreover, we have developed a simple method for removal in high-mannose. We confirmed that this method efficiently analyzed N-glycans.
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