2017 Fiscal Year Research-status Report
Study of cartilage exosomes released under mechanical stimulation and their application to cartilage regeneration
| Project/Area Number |
17K13013
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| Research Institution | The University of Tokyo |
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Principal Investigator |
モンターニュ ケヴィン 東京大学, 大学院工学系研究科(工学部), 特任講師 (50606118)
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| Project Period (FY) |
2017-04-01 – 2019-03-31
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| Keywords | exosomes / cartilage / cell signalling / hydrostatic pressure |
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| Outline of Annual Research Achievements |
Exosomes are small vesicles released by most cells and have recently attracted much focus as a very promising tool for regenerative medicine. Those vesicles have been shown to contain various proteins, messenger RNAs and micro-RNAs, and play a crucial role in cell signaling. Hydrostatic pressure (HP) is one of the main mechanical stimuli sensed by cartilage cells during joint loading in vivo and is known to affect the differentiation of cartilage, moderate pressure promoting differentiation, and excessive pressure leading to de-differentiation.
The purpose of the project is to understand how HP affects the production of cartilage exosomes and to test whether those findings can be applied to tissue engineering and the prevention of cartilage degradation in osteoarthritis.
During the first year of the project, we have optimized the method for extracting exosomes by a precipitation method, and have started the characterization of the isolated exosomes; we have shown notably that HP applied for up to 24h does not affect the quantity of exosomes released in the medium by ATDC5 cells. RNA was then extracted and characterization of the RNA species contained in the isolated exosomes is now under way.
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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
The objective of the first year was to produce and purify exosomes from cartilage cells under physiological and excessive hydrostatic pressure and start their characterization. As originally expected, we have managed to purify exosomes from the culture medium of cells submitted to high hydrostatic pressure. Thoses were quantified and it was shown that pressure applied for up to 24h did not affect their quantity. RNAs have been extracted and the identification of the RNAs contained in the exosomes is under way.
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| Strategy for Future Research Activity |
The objective of the coming year is to evaluate the efficiency of using exosomes produced under pressure to promote cartilage maturation in vitro.
To this end, the effect of exosomes produced by cartilage cells under pressure on the differentiation of engineered cartilage tissue will be investigated. Tissue constructs made from cartilage presursor cells will be cultured with exosomes produced by differentiated cartilage cells and cells under pressure. Gene and protein expression analysis will be performed to assess the differentiation status of the tissue.
The next step will be to use exosomes under excessive pressure to investigate whether such exosomes can mimic the effects of osteoarthritis on cartilage cells.
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