研究課題/領域番号 |
22H03934
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研究種目 |
基盤研究(B)
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配分区分 | 補助金 |
応募区分 | 一般 |
審査区分 |
小区分90110:生体医工学関連
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研究機関 | 東京大学 |
研究代表者 |
LECLERC Eric 東京大学, 大学院工学系研究科(工学部), 外国人客員研究員 (30759436)
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研究分担者 |
酒井 康行 東京大学, 大学院工学系研究科(工学部), 教授 (00235128)
DANOY MATHIEU 東京大学, 大学院工学系研究科(工学部), 助教 (90882621)
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研究期間 (年度) |
2022-04-01 – 2025-03-31
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研究課題ステータス |
交付 (2023年度)
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配分額 *注記 |
17,290千円 (直接経費: 13,300千円、間接経費: 3,990千円)
2023年度: 8,970千円 (直接経費: 6,900千円、間接経費: 2,070千円)
2022年度: 4,940千円 (直接経費: 3,800千円、間接経費: 1,140千円)
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キーワード | liver disease model / stem cells / organ on chip / fibrosis / liver / liver on chip / pluripotent stem cells / disease model / multi cellular cultures / MicroPhysiologicalSystem / Liver / iPSCs |
研究開始時の研究の概要 |
肝線維症は、肝がんの温床となる慢性疾患であり、先進国では患者数が急増しています。病気のプロセスを再現するヒト肝組織培養モデルがないことが、新薬や治療法の開発の障害になっています。様々な細胞種が複雑に関与する肝線維化を再現するためには、肝臓を構成する多能性細胞集団を3次元的に組織化し、血流に沿った代謝ゾーニングを再現する必要があり、線維化の発生・拡大の空間的再現に不可欠である。 本研究は、まずヒト多能性幹細胞から肝臓の4つの構成細胞を誘導し、マイクロ流体デバイス技術を応用して生理的微小培養系(マイクロフィジックスシステム;MPS)を構築し、肝線維化の進行を追跡・観察できる新規培養系を確立しました。
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研究実績の概要 |
We are planning to use organ on chip and hepatocyte like cells (HLCs), liver sinusoidal like cells (LSECs) and stellate like cells (HSCs) triple coculture as a liver fibrosis model. The fibrosis will be triggered either by palmitic acid to access physio pathological progression from steatosis to liver fibrosis; either directly by the TGF beta that is a pro fibrotic factor inducing stellate cells activation. We published a paper dealing with HLCs and LSECs cocultures. The paper focuses on the characterisation of the coculture inside the biochips in the healthy condition. We established that the biochip is relevant model to express important liver function related to zonation. Then we extended our characterisation using the three cultures in healthy condition as well. The triple culture seems to show an important development of the HSC in biochip that we are now analysing. In parallel single cells sequencing analysis were performed on preliminary tri culture experiments to extract the cell subpopulation of HLCs, LSECs, HSCs in biochips. We also include control conditions and conditions exposed to TGF beta.
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現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
In the current status, we have performed the differentiation of the induced pluripotent stem cells into hepatocyte like cells (HLCs). Then the HLCS were cultivated in a liver biochip. Then, we exposed the HLCs to palmitic acid in order to trigger lipid accumulation. Physiopathological characterisation was performed at the functional and transcriptomic levels. A publication is in preparation. this will complete preliminary data obtained with TGF beta. In parallel we have generated liver sinusoidal endothelial cells (LSEC) derived induced pluripotent stem cells and we are reproducing hepatic stellate like cells (HSCs) differentiation to be able to set up tri co cultures. We previously generated first test of tri culture in biochip and the analysis of the data are in process. We are preparing a manuscript on this data set
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今後の研究の推進方策 |
The first next step is to perform tri culture of HLCs, LSEC and HSCs to establish stable tri culture model inside the biochips.Preliminary experiment show a potential HSCs over growth that is not expected. For that purpose we need to establish the best optimise protocol of cell seeding to get healthy and uniform culture. The second next step will be to expose the tri culture to palmitic acid rather than TGF beta. The purpose is to generate a physiological development of the fibrosis using the fat accumulation as a source of stress. In parallel we are planning to analyse the preliminary TGF beta exposure that was use to simulate the fibrosis induction.
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