Experiment Overview
| Repository ID: | FR-FCM-ZZTK | Experiment name: | Fibroblast surface markers | MIFlowCyt score: | 75.22% |
| Primary researcher: | Zuzana Kahounová | PI/manager: | Karel Soucek | Uploaded by: | Zuzana Kahounová |
| Experiment dates: | 2014-12-01 - 2017-01-31 | Dataset uploaded: | Aug 2016 | Last updated: | Apr 2017 |
| Keywords: | [anti-fibroblast] [fibroblast activation protein α] [fibroblast surface protein] [epithelial-to-mesenchymal transition] [cancer-associated fibroblasts] | Manuscripts: | [28383825] |
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| Organizations: |
Institute of Biophysics AS CR, v.v.i., Department of Cytokinetics, Brno, (Czech Republic)
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| Purpose: | The purpose of this experiment was to use comercially available antibodies detecting surface markers of fibroblasts and verify their specificity on the panel of human fibroblast cells and also cells of epithelial origin. Namely, anti-FAP antibody, anti-FSP antibody and anti-fibroblast antibody were tested. | ||||
| Conclusion: | Obtained results indicate that none of tested markers was expressed only on fibroblasts, but variable expression on cells of epithelial origin was detected. Therefore, combination of markers for both positive and negative selection of fibroblast is adviced in order to identify fibroblasts unequivocaly. | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Cell debris, doublets, and aggregates were excluded from analysis based on a dual-parameter dot plot in which the pulse ratio (signal area/signal high; y-axis) versus signal area (x-axis) was displayed. Dead cells were excluded from analyses using LIVE/DEAD® Fixable Dead Cell Stain Kit. The automatic compensation for multicolor immunofluorescence analysis was performed using UltraComp eBeads (eBioscience), Anti-mouse IgG kappa/negative control (FBS) compensation particles set (BD Biosciences), and ArC Amine Reactive Compensation Bead Kit (Thermo Fisher Scientific). Compensation matrix was calculated either using FlowJo or FACSuite software (BD Biosciences), or manually corrected. | ||||
Experiment variables
| Sample Type | |
|---|---|
| · PY005 | PY005 FAP.fcs · PY005 FSP.fcs · PY005 af488.fcs · PY005 anti-fib+CD24.fcs · PY005 iso rea+mouse BV421.fcs |
| · 1329 | 1329 nontumor FAP.fcs · 1329 nontumor FSP.fcs · 1329 nontumor iso.fcs · 1329 tumor FAP.fcs · 1329 tumor FSP.fcs · 1329 tumor iso.fcs |
