演題概要

1P-28

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データ非依存的取得LC/MS/MSを用いたヒト人工多能性幹細胞由来の神経幹細胞における細胞外分泌タンパク質の解析

(横市大院生命医)
^o三上芽生・ 河内悠華子・ 高倉大輔・ 川崎ナナ

Induced pluripotent stem cell (iPSC) derived products have potential applications in regenerative medicine. For their clinical application, appropriate and efficient quality control during the manufacturing process is important. Currently, cell quality control is mainly based on the expression of intracellular molecular markers, which requires consumption of cell samples and complex manipulations. For efficient quality control, extracellular secretory proteins are useful. However, analysis of extracellular secretory proteins in culture supernatants is difficult. This is because extracellular secretory proteins are small amounts and are difficult to distinguish from proteins derived from medium. Recently, extracellular vesicles (EVs) have attracted attention as a non-invasive biomarker. EV enrichment enables distinguish from proteins derived from medium. Data independent acquisition mass spectrometry (DIA-MS) is suitable for the analysis of small amounts of proteins. Therefore, we developed a method to analyze extracellular secretory proteins focusing on extracellular vesicles by DIA-MS. Using this method, we identified extracellular secretory proteins specific to neural stem cells (NSCs) derived from human iPSCs. Many of them are involved in axon guidance and Wnt signaling pathway, reflecting the cellular state of NSCs.