研究実績の概要 |
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 was 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 shown that HP applied for up to 24h does not affect the quantity of exosomes released in the medium by ATDC5 cells. During the second year, the effects of exosomes isolated from differentiated ATDC5 cells on un-differentiated ATDC5 cells were investigated. Furthermore, RNA was extracted from exosomes for characterization bu microarray technology. During the third year, the micro-RNAs present in the isolated exosomes were identified by microarray and the microarray results are now under verification.
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