Experiment Overview
| Repository ID: | FR-FCM-Z2HL | Experiment name: | Coupling microscopy and flow cytometry for a comprehensive characterization of nanoparticle production in insect cells | MIFlowCyt score: | 54.25% |
| Primary researcher: | Eduard Puente-Massaguer | PI/manager: | Eduard Puente-Massaguer | Uploaded by: | Eduard Puente-Massaguer |
| Experiment dates: | 2018-09-01 - 2019-02-01 | Dataset uploaded: | Mar 2020 | Last updated: | Apr 2020 |
| Keywords: | [flow cytometry] [Virus-like particle] [Baculovirus] [Cryo-TEM] [Flow virometry] [Occlusion-derived virus] | Manuscripts: | [32515126] |
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| Organizations: |
Verity Software House, Verity Software House, Topsham, ME (United States)
Universitat Autònoma de Barcelona, Departament d'Enginyeria Química, Biològica i Ambiental, Cerdanyola del Vallès, Barcelona (Spain) |
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| Purpose: | Assess and compare the Gag-eGFP VLP production process in the Sf9 and High Five insect cell lines by AcMNPV baculovirus infection. | ||||
| Conclusion: | The AcMNPV baculovirus infection process progresses faster in Sf9 cells but High Five cells display a marked increase in their complexity (SSC-H), which is not observed in the former. This increase in cell complexity as the baculovirus infection advances is attributed to the production of higher levels of occlusion-derived baculoviruses in High Five over Sf9 cells. A higher concentration of Gag-eGFP VLPs is achieved in Sf9 cells although High Five cells produce larger amounts of the Gag-eGFP polyprotein. Moreover, the interaction of High Five-derived VLPs with cellular compounds is detected, which increases the sedimentation coefficient of these nanoparticles | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | DPBS and fresh Sf900III medium were used as controls. Furthermore, non-infected sample supernatants (0 hours post infection) and detergent lysis controls were also included. All experiments were performed in triplicate | ||||
Experiment variables
| Individuals | |
|---|---|
| · Cells | High Five cells - 0 hpi.fcs · High Five cells - 16 hpi.fcs · High Five cells - 24 hpi.fcs · High Five cells - 40 hpi.fcs · Sf9 cells - 0 hpi.fcs · Sf9 cells - 16 hpi.fcs · Sf9 cells - 24 hpi.fcs · Sf9 cells - 40 hpi.fcs |
| · Nanoparticles | Fresh DPBS control.fcs · High Five nanoparticles - 0 hpi.fcs · High Five nanoparticles - 16 hpi.fcs · High Five nanoparticles - 24 hpi.fcs · High Five nanoparticles - 40 hpi.fcs · Sf9 nanoparticles - 0 hpi.fcs · Sf9 nanoparticles - 16 hpi.fcs · Sf9 nanoparticles - 24 hpi.fcs · Sf9 nanoparticles - 40 hpi.fcs · High Five nanoparticles - detergent lysis control.fcs · Sf9 nanoparticles - detergent lysis control.fcs · Fresh Sf900III medium control.fcs |
