| 研究実績の概要 |
1) We acquired a library of 6 ALS and 4 healthy control human iPSC lines from Cedar Sinai Cell Bank. 2) We established a protocol of differentiation of motor neurons (MNs) from these human IPSC lines. 3) We characterized axonal degeneration, protein aggregation and neuronal cell death in our human MNs cultures. 4) We looked at the role of mitochondria and ATP production in correlation with local translation and phase separation for all the lines. 5) We collected cell pellets and culture medium from pre-symptomatic, early onset and late stage of disease MNs in culture. 6) We optimized a protocol for isolation of extracellular vesicles (EVs).
FINAL YEAR
7) We characterized the retrieved EVs biochemically and by electron microscopy. 8) We optimized the condition from proteomics analysis of MN culture media and extracted EVs. 9) We performed proteomics with the samples described in 3-4 to identify new ALS biomarkers. 10) We conducted a bioinformatic analysis to compare the different cell lines for each time point. 11) We are proceeding with candidate biomarker validation both in culture and from patient fluids.
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