Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasis
Abstract Background The recruitment of effector cells is one of the novel functions described for extracellular vesicles (EVs) that needs further study. For instance, cell recruitment by mesenchymal stromal cell derived-EVs (MSC-EVs) is one of the features by which MSC-EVs may induce regeneration an...
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| Format: | Article |
| Language: | English |
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BMC
2023-10-01
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| Series: | BMC Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12915-023-01729-5 |
| _version_ | 1797647183766355968 |
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| author | Marta Clos-Sansalvador Marta Monguió-Tortajada Ferran Grau-Leal Vicenç Ruiz de Porras Sergio G. Garcia Marta Sanroque-Muñoz Miriam Font-Morón Marcella Franquesa Francesc E. Borràs |
| author_facet | Marta Clos-Sansalvador Marta Monguió-Tortajada Ferran Grau-Leal Vicenç Ruiz de Porras Sergio G. Garcia Marta Sanroque-Muñoz Miriam Font-Morón Marcella Franquesa Francesc E. Borràs |
| author_sort | Marta Clos-Sansalvador |
| collection | DOAJ |
| description | Abstract Background The recruitment of effector cells is one of the novel functions described for extracellular vesicles (EVs) that needs further study. For instance, cell recruitment by mesenchymal stromal cell derived-EVs (MSC-EVs) is one of the features by which MSC-EVs may induce regeneration and ameliorate tissue injury. On the other hand, increasing evidence suggests that cancer EVs play an important role in the preparation of the pre-metastatic niche (PMN) by recruiting their primary tumour cells. Understanding and measuring the potential of MSC-EVs or cancer-EVs to induce cell migration and recruitment is essential for cell-free therapeutic approaches and/or for a better knowledge of cancer metastasis, respectively. In this context, classical in vitro migration assays do not completely mimic the potential situation by which EVs exert their chemotactic capacity. Results We adapted an agarose spot migration assay as an in vitro system to evaluate the cell recruitment capacity of locally delivered or localized EVs. Cell migration was tracked for 12 h or 48 h, respectively. Thereafter, endpoint migration images and time-lapse videos were analysed to quantify several parameters aiming to determine the migration of cells to either MSC-EV or pro-metastatic EV. The number of cells contained inside the agarose spots, the migration distance, the area occupied by cells, the directionality of the cell movement, and the Euclidean distance were measured. This multi-parametric evaluation revealed the potential of different MSC-EV preparations to recruit endothelial cells and to detect an enhanced recruitment capacity of highly metastatic PC3-derived EVs (PC3-EVs) compared to low-metastatic LNCaP-EVs in a tumour cell-specific manner. Conclusions Overall, this agarose spot migration assay may offer a diversity of measurements and migration settings not provided by classical migration assays and reveal its potential use in the EV field in two different contexts with recruitment in common: regeneration and cancer metastasis. |
| first_indexed | 2024-03-11T15:12:38Z |
| format | Article |
| id | doaj.art-49b7b514e3094bbbb69006ba19849e59 |
| institution | Directory Open Access Journal |
| issn | 1741-7007 |
| language | English |
| last_indexed | 2024-03-11T15:12:38Z |
| publishDate | 2023-10-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biology |
| spelling | doaj.art-49b7b514e3094bbbb69006ba19849e592023-10-29T12:36:33ZengBMCBMC Biology1741-70072023-10-0121111110.1186/s12915-023-01729-5Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasisMarta Clos-Sansalvador0Marta Monguió-Tortajada1Ferran Grau-Leal2Vicenç Ruiz de Porras3Sergio G. Garcia4Marta Sanroque-Muñoz5Miriam Font-Morón6Marcella Franquesa7Francesc E. Borràs8REMAR-IGTP Group, Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP)REMAR-IGTP Group, Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP)RCPB Group, CARE Program, Germans Trias i Pujol Research Institute (IGTP); ProCURE Program, Catalan Institute of Oncology, Carretera de Can RutiCARE Program, Germans Trias i Pujol Research Institute (IGTP)REMAR-IGTP Group, Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP)REMAR-IGTP Group, Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP)REMAR-IGTP Group, Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP)REMAR-IGTP Group, Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP)REMAR-IGTP Group, Germans Trias i Pujol Research Institute (IGTP) & Nephrology Department, University Hospital Germans Trias i Pujol (HUGTiP)Abstract Background The recruitment of effector cells is one of the novel functions described for extracellular vesicles (EVs) that needs further study. For instance, cell recruitment by mesenchymal stromal cell derived-EVs (MSC-EVs) is one of the features by which MSC-EVs may induce regeneration and ameliorate tissue injury. On the other hand, increasing evidence suggests that cancer EVs play an important role in the preparation of the pre-metastatic niche (PMN) by recruiting their primary tumour cells. Understanding and measuring the potential of MSC-EVs or cancer-EVs to induce cell migration and recruitment is essential for cell-free therapeutic approaches and/or for a better knowledge of cancer metastasis, respectively. In this context, classical in vitro migration assays do not completely mimic the potential situation by which EVs exert their chemotactic capacity. Results We adapted an agarose spot migration assay as an in vitro system to evaluate the cell recruitment capacity of locally delivered or localized EVs. Cell migration was tracked for 12 h or 48 h, respectively. Thereafter, endpoint migration images and time-lapse videos were analysed to quantify several parameters aiming to determine the migration of cells to either MSC-EV or pro-metastatic EV. The number of cells contained inside the agarose spots, the migration distance, the area occupied by cells, the directionality of the cell movement, and the Euclidean distance were measured. This multi-parametric evaluation revealed the potential of different MSC-EV preparations to recruit endothelial cells and to detect an enhanced recruitment capacity of highly metastatic PC3-derived EVs (PC3-EVs) compared to low-metastatic LNCaP-EVs in a tumour cell-specific manner. Conclusions Overall, this agarose spot migration assay may offer a diversity of measurements and migration settings not provided by classical migration assays and reveal its potential use in the EV field in two different contexts with recruitment in common: regeneration and cancer metastasis.https://doi.org/10.1186/s12915-023-01729-5Extracellular vesiclesExosomesMigrationRecruitmentAgaroseCancer |
| spellingShingle | Marta Clos-Sansalvador Marta Monguió-Tortajada Ferran Grau-Leal Vicenç Ruiz de Porras Sergio G. Garcia Marta Sanroque-Muñoz Miriam Font-Morón Marcella Franquesa Francesc E. Borràs Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasis BMC Biology Extracellular vesicles Exosomes Migration Recruitment Agarose Cancer |
| title | Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasis |
| title_full | Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasis |
| title_fullStr | Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasis |
| title_full_unstemmed | Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasis |
| title_short | Agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles: applications in regenerative medicine and cancer metastasis |
| title_sort | agarose spot migration assay to measure the chemoattractant potential of extracellular vesicles applications in regenerative medicine and cancer metastasis |
| topic | Extracellular vesicles Exosomes Migration Recruitment Agarose Cancer |
| url | https://doi.org/10.1186/s12915-023-01729-5 |
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