Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles
Summary: Hematopoietic stem cells (HSCs) maintain their self-renewal capacity in an autocrine manner through hematopoietic stem and progenitor cell (HSPC)-derived extracellular vesicles (EVs). Here, we present a protocol for the isolation and characterization of EVs from HSPCs starting from an in vi...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-06-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725001844 |
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| author | Federica Zanotti Ilaria Zanolla Massimo Bonora Claudia Morganti Barbara Zavan Letizia Ferroni Paolo Pinton Keisuke Ito |
| author_facet | Federica Zanotti Ilaria Zanolla Massimo Bonora Claudia Morganti Barbara Zavan Letizia Ferroni Paolo Pinton Keisuke Ito |
| author_sort | Federica Zanotti |
| collection | DOAJ |
| description | Summary: Hematopoietic stem cells (HSCs) maintain their self-renewal capacity in an autocrine manner through hematopoietic stem and progenitor cell (HSPC)-derived extracellular vesicles (EVs). Here, we present a protocol for the isolation and characterization of EVs from HSPCs starting from an in vivo murine model. We describe steps for murine bone marrow isolation, HSPC staining and sorting, HSPC-derived EV isolation, size and concentration characterization, and EV visualization and marker description.For complete details on the use and execution of this protocol, please refer to Bonora et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-51bdb98166b14a918afbf78ee304ac6a |
| institution | DOAJ |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-51bdb98166b14a918afbf78ee304ac6a2025-08-20T03:18:19ZengElsevierSTAR Protocols2666-16672025-06-016210377810.1016/j.xpro.2025.103778Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesiclesFederica Zanotti0Ilaria Zanolla1Massimo Bonora2Claudia Morganti3Barbara Zavan4Letizia Ferroni5Paolo Pinton6Keisuke Ito7Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Departments of Oncology and Medicine, Albert Einstein College of Medicine-Montefiore Health System, Bronx, NY 10461, USADepartment of Medical Sciences, University of Ferrara, 44121 Ferrara, ItalyDepartment of Medical Sciences, University of Ferrara, 44121 Ferrara, ItalyRuth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Departments of Oncology and Medicine, Albert Einstein College of Medicine-Montefiore Health System, Bronx, NY 10461, USAMaria Cecilia Hospital, GVM Care & Research, Cotignola, 48033 Ravenna, Italy; Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, 44121 Ferrara, Italy; Translational Medicine Department, University of Ferrara, 44121 Ferrara, ItalyMaria Cecilia Hospital, GVM Care & Research, Cotignola, 48033 Ravenna, ItalyDepartment of Medical Sciences, University of Ferrara, 44121 Ferrara, ItalyRuth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Departments of Oncology and Medicine, Albert Einstein College of Medicine-Montefiore Health System, Bronx, NY 10461, USA; Corresponding authorSummary: Hematopoietic stem cells (HSCs) maintain their self-renewal capacity in an autocrine manner through hematopoietic stem and progenitor cell (HSPC)-derived extracellular vesicles (EVs). Here, we present a protocol for the isolation and characterization of EVs from HSPCs starting from an in vivo murine model. We describe steps for murine bone marrow isolation, HSPC staining and sorting, HSPC-derived EV isolation, size and concentration characterization, and EV visualization and marker description.For complete details on the use and execution of this protocol, please refer to Bonora et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725001844Cell BiologyCell isolationFlow CytometryMetabolismStem Cells |
| spellingShingle | Federica Zanotti Ilaria Zanolla Massimo Bonora Claudia Morganti Barbara Zavan Letizia Ferroni Paolo Pinton Keisuke Ito Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles STAR Protocols Cell Biology Cell isolation Flow Cytometry Metabolism Stem Cells |
| title | Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles |
| title_full | Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles |
| title_fullStr | Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles |
| title_full_unstemmed | Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles |
| title_short | Protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell-derived extracellular vesicles |
| title_sort | protocol for the isolation and characterization of murine hematopoietic stem and progenitor cell derived extracellular vesicles |
| topic | Cell Biology Cell isolation Flow Cytometry Metabolism Stem Cells |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725001844 |
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