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<p>We propose an oil-sealed, arginine-glycine-aspartic (RGD) -modified alginate hydrogel microwell array for the analysis of single-cell-derived extracellular vesicles and particles (EVPs) secreted by adherent cells cultured in enclosed spaces. Taking advantage of the mesh structure of alginate hydrogel, we developed a microwell array with size-selective permeability that allows nutrients to pass through the hydrogel while preventing EVPs from doing so. Continuous single-cell culture in sealed microwells (&gt;19 days) has been achieved, while retaining the EVPs inside the microwells. The single-cell-derived EVPs were collected from sealed microwells using a glass capillary to analyze surface membrane proteins. We believe that our oil-sealed RGD-modified alginate hydrogel microwell array will contribute to revealing the heterogeneity of cells, thereby advancing our understanding of the mechanisms of various diseases.</p>

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Oil-sealed RGD-modified alginate hydrogel microwell array for analysis of single-cell-derived extracellular vesicles and particles

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年12月2日

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  • Chisaki Yamagata,
  • Yuto Hamazaki,
  • Tomoharu Nakazato,
  • Shun Itai,
  • Mai Honjo,
  • Mami Kato,
  • Yuta Kurashina,
  • Makoto Asai,
  • Ayuko Hoshino,
  • Hiroaki Onoe

摘要

We propose an oil-sealed, arginine-glycine-aspartic (RGD) -modified alginate hydrogel microwell array for the analysis of single-cell-derived extracellular vesicles and particles (EVPs) secreted by adherent cells cultured in enclosed spaces. Taking advantage of the mesh structure of alginate hydrogel, we developed a microwell array with size-selective permeability that allows nutrients to pass through the hydrogel while preventing EVPs from doing so. Continuous single-cell culture in sealed microwells (>19 days) has been achieved, while retaining the EVPs inside the microwells. The single-cell-derived EVPs were collected from sealed microwells using a glass capillary to analyze surface membrane proteins. We believe that our oil-sealed RGD-modified alginate hydrogel microwell array will contribute to revealing the heterogeneity of cells, thereby advancing our understanding of the mechanisms of various diseases.