Isolation of Exosome-Enriched Extracellular Vesicles Carrying Granulocyte-Macrophage Colony-Stimulating Factor from Embryonic Stem Cells
| Autor: | Aaron G. Whitt, Kavitha Yaddanapudi, Shuhan Meng, Allison Tu, Chi Li, John W. Eaton |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
General Immunology and Microbiology
Chemistry General Chemical Engineering General Neuroscience Granulocyte-Macrophage Colony-Stimulating Factor Cell Differentiation Extracellular vesicle Exosomes Embryonic stem cell Exosome Article General Biochemistry Genetics and Molecular Biology Microvesicles Cell biology Mice Granulocyte macrophage colony-stimulating factor Cell culture Extracellular medicine Animals Cytokines Induced pluripotent stem cell Embryonic Stem Cells medicine.drug |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| DOI: | 10.3791/60170 |
| Popis: | Embryonic stem cells (ESCs) are pluripotent stem cells capable of self-renewal and differentiation into all types of embryonic cells. Like many other cell types, ESCs release small membrane vesicles, such as exosomes, to the extracellular environment. Exosomes serve as essential mediators of intercellular communication and play a basic role in many (patho)physiological processes. Granulocyte-macrophage colony-stimulating factor (GM-CSF) functions as a cytokine to modulate the immune response. The presence of GM-CSF in exosomes has the potential to boost their immune regulatory function. Here, GM-CSF was stably overexpressed in the murine ESC cell line ES-D3. A protocol was developed to isolate high-quality exosome-enriched extracellular vesicles (EVs) from ES-D3 cells overexpressing GM-CSF. Isolated exosome-enriched EVs were characterized by a variety of experimental approaches. Importantly, significant amounts of GM-CSF were found to be present in exosome-enriched EVs. Overall, GM-CSF-bearing exosome-enriched EVs from ESCs might function as cell-free vesicles to exert their immune regulatory activities. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|