本研究では生体外で細胞を三次元的に培養するための足場材料として，高静水圧を用いた脱細胞化組織の作製することを目的としている．最終的には，脱細胞化組織を用いた肝・腎組織の再生医療への適用を目指している．本年度は，前年度に開発した油圧駆動型の超高静水圧処理装置を改良して周期的に高圧と低圧の印加を与えられるようにし，脱細胞効果の向上をはかった．さらに，こちらの装置を用いて，ヒト皮膚由来の細細胞に対して高静水圧の繰り返し印加を実施して，脱細胞化に関わる因子である脱核および細胞死に与える影響を評価した．その結果，細胞懸濁液を対象とした試験では250 MPa，5回程度繰り返し印加することで，一般的に超高静水圧で適用される1000 MPaと同程度の細胞死を誘導し，さらに脱核が実現できることを明らかにした．今後は細胞とゲルからなる三次元培養体やラット腎組織などの動物組織を対象として繰り返し高静水圧処理を用いた脱細胞化組織の作製を実施し，高静水圧の印加条件が細胞死，脱核に与える影響を明らかにする．
The purpose of this study is to produce decellularized tissue using high hydrostatic pressure as a scaffold for in vitro three-dimensional cell culture. Final goal of this study is to apply this decellularized material for liver and kidney tissue regeneration. This year, we improved the hydraulically-driven ultra-high hydrostatic pressure treatment device for cyclic application to increase the decellularization effect. Furthermore, using this device, human skin-derived cell suspension were subjected to the cyclic ultra-high hydrostatic pressure and the effects on enucleation and cell death, which are factors involved in decellularization, were evaluated. As a result, by cyclic application of 250 MPa about 5 times, cell death was induced to the same extent as application of 1000 MPa in static condition, which is a general condition for decellularization. Furthermore, the enucleation was also observed. In the future, we plan to examine the decellularization effect for three-dimensional tissues such as three-dimensional cultures consisting of cells and hydro gels and rat kidney tissues