Institute of Nano-Life-Systems, Institutes of Innovation for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Sci Rep. 2021 Apr 21;11(1):8672. doi: 10.1038/s41598-021-87986-2.
Recent studies have shown that extracellular vesicles (EVs) can be utilized as appropriate and highly specific biomarkers in liquid biopsy for the diagnosis and prognosis of serious illness. However, there are few methods that can collect and isolate miRNA in EVs simply, quickly and efficiently using general equipment such as a normal centrifuge. In this paper, we developed an advanced glass membrane column (AGC) device incorporating a size-controlled macro-porous glass (MPG) membrane with a co-continuous structure to overcome the limitations of conventional EV collection and miRNA extraction from the EVs. The size of macro-pores in the MPG membrane could be accurately controlled by changing the heating temperature and time on the basis of spinodal decomposition of BO, NaO, and SiO in phase separation. The AGC device with an MPG membrane could collect the EVs simply and quickly (< 10 min) from cell culture supernatant, serum and urine. This AGC device could extract miRNA from the EVs captured in the MPG membrane with high efficiency when combined with a miRNA extraction solution. We suggest that the AGC device with an MPG membrane can be useful for the diagnosis and prognosis of serious illness using of EVs in various kinds of body fluids.
最近的研究表明，细胞外囊泡 (EVs) 可以作为适当的、高度特异性的生物标志物，用于液体活检中严重疾病的诊断和预后。然而，很少有方法可以使用普通设备（如普通离心机）简单、快速和有效地收集和分离 EV 中的 miRNA。在本文中，我们开发了一种先进的玻璃膜柱 (AGC) 装置，该装置结合了具有共连续结构的尺寸可控的大孔玻璃 (MPG) 膜，以克服传统 EV 收集和从 EV 中提取 miRNA 的局限性。通过改变加热温度和时间，可以在 BO、NaO 和 SiO 相分离的旋节分解的基础上，精确控制 MPG 膜中大孔的尺寸。具有 MPG 膜的 AGC 装置可以简单快速地(<10 min)从细胞培养上清液、血清和尿液中收集 EV。当与 miRNA 提取溶液结合使用时，该 AGC 装置可以从 MPG 膜中捕获的 EV 中高效提取 miRNA。我们认为，具有 MPG 膜的 AGC 装置可用于使用各种体液中的 EV 进行严重疾病的诊断和预后。
