成果報告書詳細
管理番号20130000000684
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(国立大学法人大阪大学蛋白質研究所)
公開日2014/8/20
報告書年度2012 - 2012
委託先名国立大学法人大阪大学蛋白質研究所
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Title: The development to accelerate the industrial application of human stem cells.
The development of basic evaluation technology for the industrialization of human stem cells (FY2010-FY2015) FY2012Annual Report

(1) Research item 1: The development of the technology for stable cultivation and preservation of human stem cells
1-1 The development of an automatic cell processing and freezing preservation machine for the stable cultivation
Using the automatic cell culturing system, that is a platform for the development, installed at Kyoto Laboratory, an automatic freezing and thawing of iPS cells established by Kyoto Univ. CiRA has been implemented. Using the cooperation function, the automatic culture iPS cells thawed automatically has become possible.
The freezing and thawing functions was improved. As a result, the trial cryopreservation machine linked the auto-culture machine was completed.
In the culture of human iPS cells using SNL, we began the set of automated processing evaluation test about pre-freezing, cryopreservation, thawing and re-cultured.
For the cell observation device, the software for noninvasive estimation of human iPS cell quality and characteristics has been developed and validated. This software enables to determine the differentiated region using morphological parameters measured by the shape of the boundaries, image contrast pattern, size and compactness of the culturing iPS colonies. The first proto-type of minimally invasive 3-D imaging has been developed.

1-2 The development of the culture substrates and medium optimized for stable cultivation of stem cells
For the culture substrates, two prototypes of the functionally improved laminin 511 fragments have been produced and their multifunctionality has been confirmed. The stability of the laminin 511 fragment to drying after coating onto culture vessels has been assessed. Candidate molecules have been screened for their ability to protect the laminin fragment from inactivation after drying.
For the medium, effective several candidate culture medium have been selected through the evaluation on the affinity with the laminin 511. General cell stocks of CiRA iPS lines have been developed to be able to perform more stable evaluations of devices through the project.
By using the commercial freezing medium for embryonic stem cells, it confirmed that the automatic freezing preservation and thawing of human iPS cells were possible. As a result, the simple freezing preservation of human iPS cells by automatic freezing / thawing machine became possible.

1-3.The development of the technology for quality control and table cultivation of mesenchymal stem cells
Integrated cellular data have been acquired based on cellular proliferation and differentiation ability of various types of MSCs using either serum or serum-free medium. Global gene expression analysis and lectin micro array analysis for these MSCs samples have been done and various specific markers have been identified. Particularly, novel biomarkers suitable for prolferation ability and stem cell multipotency have been identified

(2) Research item 2: The development of the technology of quality control and stable cultivation of human stem cells
2-1 Proposal of draft standards of technologies around human iPS cells
Organized and held a study debate meeting entitled by "Standardization of terms and definitions in the field of regenerative medicine and stem cell technology," to which participants include experts from the Japanese society of regenerative medicine, universities in the field of tissue engineering, Japanese biotech industries and other stakeholders from Japanese government board and ministries, resulting in a draft document of ISO International Standards for ISO/TC 276 Biotechnology on biotech terminology or terms and definitions of human stem cell types and related processing and production.
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