Experiment Overview
| Repository ID: | FR-FCM-Z32H | Experiment name: | vFC of THP-1 EVs | MIFlowCyt score: | 63.25% |
| Primary researcher: | John Nolan | PI/manager: | John Nolan | Uploaded by: | John Nolan |
| Experiment dates: | 2019-01-01 - 2020-06-30 | Dataset uploaded: | Oct 2020 | Last updated: | Oct 2020 |
| Keywords: | [CD63] [CD9] [exosome] [extracellular vesicle] [antigen-presenting cells] [THP-1] [luciferase] | Manuscripts: | [33935791] [PMC8085554] | ||
| Organizations: |
Scintillon Institute, San Diego, CA (USA)
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| Purpose: | Characterization of extracellular vesicles released by THP-1 cell using vesicle flow cytometry (vFC). To enable drug discovery and characterization of compounds that affect EV biogenesis, function, and release, we developed and characterized a reporter cell line that allows the quantitation of EV shed into culture media. | ||||
| Conclusion: | These results indicate that the use of CD63Tluc-CD9EmGFP reporter cells is feasible for HTS of compounds that regulate EV release. | ||||
| Comments: | None | ||||
| Funding: | This work was supported by the National Institute of Health/ National Institute of Allergy and Infectious Diseases under contracts HHSN272201400051C and 75N93019C00042. CL was supported by the Medical Scientific Research Foundation of Guangdong Province, China (2018037), and the National Natural Science Foundation of China (81503215). | ||||
| Quality control: | Instrument performance was characterized using a combination of multi-intensity single fluorophore beads (Quantum FITC, Bangs Labs Quantibrite PE, BD Biosciences) whose intensity had been calibrated in units of MESF, multi-intensity multifluorophore beads (vCal nanoRainbow, Cellarcus), and antibody capture beads (vCal antibody capture beads, Cellarcus) calibrated to report results in units of antibodies bound per vesicle (ABV). EV analysis by vesicle flow cytometry (VFC) was conducted and reported as suggested by the MIFlowCyt-EV guidelines (See attached checklist). | ||||
