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 ...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.展开更多
基金supported by Research Project Keio 2040(Creativity Initiative)at Keio University Global Research Institute,JapanJST SPRING(JPMJSP2123 to C.Y.)+3 种基金JSPS Grant-in-Aid for JSPS Research Fellow(24KJ0984 to Y.H.,24KJ0956 to M.H.)JSPS KAKENHI(JP25KJ0103 to M.K.,JP20H05904 and JP25H01066 to A.H.)JST FOREST(JPMJFR216B to A.H.)JST Moonshot MILLENNIA Program(JPMJMS2292 to A.H.)AMED(24zf0127012 and 22zf0127009h000 to A.H.).
文摘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.
