成果報告書詳細
管理番号20130000000496
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/Muse細胞の評価基盤技術開発」
公開日2014/8/20
報告書年度2012 - 2012
委託先名国立大学法人東北大学 株式会社Clio 国立大学法人名古屋大学 国立大学法人京都大学
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Development of Basic Technology for the Promotion of Industrial Applications for Human Stem Cells / Development of evaluation technology for commercialization of human stem cells / Development of evaluation-based Muse cell technology (FY2011-FY2013) FY2012 Annual Report

In this project, we will develop indicators for the quality evaluation of Muse cells. Muse cells are non-tumorigenic pluripotent stem cells and are taken from adult human mesenchymal tissues. Moreover, we aim to establish a basis for the stable supply of a large amount of Muse cells of assured quality by developing quality evaluation indicator technology.

1. Analysis of gene/protein expression in Muse cells and development of indicators for evaluation
1-1 Analysis of the ability of suspension-cultured cells to form clusters
We have examined Muse cells isolated from the various tissues such as skin, fat and bone marrow and the isolation method examined in 2. for their ability to form clusters.

1-2 Analysis of the differentiation efficiency of adherent cultured cells into triploblastic cells
We have examined Muse cells isolated from the various tissues such as skin, fat and bone marrow and the isolation method examined in 2. for their ability to differentiate into three germ layers in adherent cultured cells.

1-3 Analysis of gene expression
We have searched for the genes specific to Muse cells by analyzing gene expression in non-Muse cells and Muse cells purified from skin, fat and bone marrow by using next-generation sequencing methods.

1-4 Proteomic analysis
In collaboration with Prof. Toshiaki Isobe of Tokyo Metropolitan University, we have performed proteomic analysis of several pairs of Muse cells and non-Muse cells by using a combined system of high-throughput liquid chromatography and tandem mass spectrometry, and have made a list of proteins specific to Muse cells.

1-5 Characterization of Muse cells by comparison with other organisms
We have analyzed the gene expression of the mesenchymal stem cells of other organisms which are equivalent to Muse cells to search for markers characteristic of those cells. In order to obtain the reference data for characterization of human Muse cells, we have searched for cells equivalent to Muse cells in mice, rats, rabbits, pigs and sheep.

1-6 In vivo examination of tissue repair ability
In collaboration with Prof. Isao Sakaida of Yamaguchi University, we have addressed the preparation of a mouse model of hepatic cirrhosis using immunodeficient mice in order to examine the tissue repair ability of Muse cells in a mouse model of hepatic cirrhosis.

2. Improvement of techniques for extraction, isolation and culture of Muse cells
To optimize the methods for isolation and culture of Muse cells, we have examined the isolation methods other than the use of a cell sorter, and also succeeded in the isolation of Muse cells from xeno-free bone marrow.

3. Test for reproducibility
Numbers of teams from home and abroad including Prof. Katsuto Tamai’s group (Osaka University) have made a presentation about the methods for the isolation and culture of Muse cells.
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