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Analysis of extracellular vesicles as a potential index for monitoring differentiation of neural lineage cells from induced pluripotent stem cells
Journal of Bioscience and Bioengineering ( IF 2.8 ) Pub Date : 2021-07-18 , DOI: 10.1016/j.jbiosc.2021.06.004
Hikaru Saito 1 , Midori Kato 1 , Kakuro Hirai 1 , Masaharu Kiyama 1 , Kunio Ohyama 1 , Hiroko Hanzawa 1 , Atsushi Nakane 2 , Sayaka Sekiya 2 , Kenji Yoshida 2 , Akiyoshi Kishino 2 , Atsushi Tsuchida 3 , Toru Kimura 3 , Jun Takahashi 4 , Shizu Takeda 1
Affiliation  

To improve cell production efficacy, it is important to evaluate cell conditions during culture. Extracellular vesicles (EVs) secreted from various cells are involved in stem cell differentiation. As EVs carry information about their source cells, we hypothesized that they may serve as a noninvasive index of cell conditions. We evaluated changes in EV morphology, concentration, and microRNA (miRNA) and protein expression in culture supernatants during the differentiation of induced pluripotent stem cells (iPSCs) into neural lineage cells, for application in regenerative medicine for Parkinson's disease. We observed EVs (50–150 nm) in culture supernatants of iPSCs and differentiated cells. The EVs expressed the exosome markers CD63, CD81, and CD9. Throughout differentiation, the EV concentration in the supernatants decreased, and EV miRNA and protein expression changed substantially. Especially, miR-106b, involved in neural stem cell differentiation and normal brain development, was considerably downregulated. CD63 expression correlated with the CORIN-positive cell rate, which is an index of differentiation. Thus, EV concentration and miRNA and protein expression may reflect the differentiation status of iPSCs. These findings pave the way for the development of novel and sensitive cell culture monitoring methods.



中文翻译:

分析细胞外囊泡作为监测诱导多能干细胞分化的神经谱系细胞的潜在指标

为了提高细胞生产效率,在培养过程中评估细胞条件非常重要。各种细胞分泌的细胞外囊泡 (EV) 参与干细胞分化。由于 EV 携带有关其源细胞的信息,我们假设它们可以作为细胞状况的非侵入性指标。我们评估了诱导多能干细胞 (iPSC) 分化为神经谱系细胞期间培养上清液中 EV 形态、浓度和 microRNA (miRNA) 和蛋白质表达的变化,以应用于帕金森病的再生医学。我们在 iPSC 和分化细胞的培养上清液中观察到 EV(50-150 nm)。EVs 表达外泌体标记 CD63、CD81 和 CD9。在整个分化过程中,上清液中的 EV 浓度下降,EV miRNA 和蛋白质表达发生了显着变化。特别是,参与神经干细胞分化和正常大脑发育的 miR-106b 显着下调。CD63 表达与作为分化指标的 CORIN 阳性细胞率相关。因此,EV 浓度和 miRNA 和蛋白质表达可能反映 iPSC 的分化状态。这些发现为开发新的和敏感的细胞培养监测方法铺平了道路。EV 浓度和 miRNA 和蛋白质表达可能反映 iPSCs 的分化状态。这些发现为开发新的和敏感的细胞培养监测方法铺平了道路。EV 浓度和 miRNA 和蛋白质表达可能反映 iPSCs 的分化状态。这些发现为开发新的和敏感的细胞培养监测方法铺平了道路。

更新日期:2021-07-18
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