2022 Fiscal Year Annual Research Report
Liver fibrosis investigation using iPSCs hepatic derived organ on chip
| Project/Area Number |
22H03934
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| Allocation Type | Single-year Grants |
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| Research Institution | The University of Tokyo |
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Principal Investigator |
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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| Co-Investigator(Kenkyū-buntansha) |
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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| Project Period (FY) |
2022-04-01 – 2025-03-31
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| Keywords | liver on chip / pluripotent stem cells / disease model / multi cellular cultures |
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| Outline of Annual Research Achievements |
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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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
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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| Strategy for Future Research Activity |
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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