成果報告書詳細
管理番号20110000001420
タイトル*平成22年度中間年報 「次世代再生医療技術の研究開発 少量の細胞により生体内で自己組織の再生を促す自律成熟型再生デバイスの開発(生体内で自律的に成熟する臓器再生デバイスのための基盤研究開発)」
公開日2011/11/23
報告書年度2010 - 2010
委託先名国立大学法人東京大学 国立大学法人大阪大学 学校法人東京理科大学 野村ユニソン株式会社 国立大学法人神戸大学 学校法人福田学園
プロジェクト番号P10004
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.研究開発の内容および成果等 本プロジェクトでは、軟骨組織再生技術の汎用化、産業化を進めるため、臓器構造体としての軟骨の自律成熟に着目し、関節やその他の欠損部位を修復させるための基盤技術を確立する。このため平成22 年度は、以下の項目の研究開発を行った。 (1)「軟骨用自律再生デバイスのための細胞培養法の開発」 (東京大学) ヒトおよびマウスの耳介軟骨細胞、関節軟骨細胞、骨髄細胞、脂肪幹細胞、滑膜細胞を用いて、ハイドロゲル(合成ペプチド、コラーゲンゲルなど)と混和し、3 次元培養を比較検討したところ、RADA を基調とした合成ペプチドやコラーゲンのゲルで耳介軟骨細胞や骨髄細胞で高い増殖能がみられた。加えて、刺激因子の最適な濃度と組み合わせを統計学的に解析し、耳介軟骨細胞や骨髄細胞において3 次元培養における高い増殖能や軟骨基質産生能を見出した。
英文要約Title: Fundamental Research Project for Autonomous Regeneration Device (FY2011-FY2013) FY2011 Annual Report
Reconstruction of joints, based on autonomous regeneration of cartilages, would enable to promote generalization and industrialization of cartilage tissue engineering. In this project, we will establish a fundamental technology to apply the autonomous regeneration for reconstruction of joints. This financial year, for the development of a cell culture technique for an autonomous regeneration device for cartilages, we adopted auricular chondrocytes, articular chondrocytes, bone marrow cells, adipose stem cells, or synoviocytes, mixed them into a hydrogel (synthetic peptide, collagen gel, etc), and attempted 3D culture. We also analyzed the optimal concentration and combination of stimulating factors statistically, and selected the cells or the hydrogels. On bones, tetrabone (tetrapod-shaped bone fillers) and β-TCP bone fillers were evaluated for the mechanical strength at 8 weeks after implantation in the canine femur defect model. The elastic modulus of TetraBone group was significant higher than β-TCP group. These results suggest that TetraBone facilitated new bone formation more efficiently, probably through better intraconnectivity of pores between Tetrabones. We also confirmed that Tetrabone had an ability to release a small molecule, which accelerates the bone differentiation, in a sustained manner. For joints, we have investigated the proliferative effects of serum-free media on mesenchymal stem cells (MSCs) derived from synovium. Serum-free media have promoted cell proliferation, compared to conventional media. As the production of hollow fibers made of biodegradable polymers, cellulose acetate butyrate hollow fiber membranes were successfully prepared. The water permeability of the obtained membrane was 650L/(m2 h atm), which exceeds the target value of 500L/(m2 h atm). The pore size of the membrane was 0.1μm. As development of a hollow fiber module, we carried out the resolvability evaluation on the hollow fiber of cellulose acetate as the biodegradability material. In addition, we compared three kinds of sterilization of autoclave / γ beam / EOG and non-sterilization product to consider influence of sterilization. The properties of matter of the hollow fiber were changed by autoclave sterilizatio, and the cellulose acetate was confirmed not to be used after this sterilization method. As exploratory animal experiment for for cartilages, we produced a culture module for transplantation, by way of trial, and examined its properties. For joints, we are developing the complexes of small particles of artificial bone which are associated each other by fibrin network. As an initial step, we optimized to develop a gel which is made of plasma from autologous peripheral blood in a short time by the optimized adjustment of the ratio of plasma to calcium chloride.
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