Experiment Overview
| Repository ID: | FR-FCM-Z3RV | Experiment name: | Tracking cell cycling activities in RPE-fucci cells | MIFlowCyt score: | 76.00% |
| Primary researcher: | Qian Wang | PI/manager: | Manuel Goncalves | Uploaded by: | Qian Wang |
| Experiment dates: | 2020-12-20 - 2021-01-08 | Dataset uploaded: | May 2021 | Last updated: | May 2021 |
| Keywords: | [cell cycle] [RPE-fucci] | Manuscripts: | [34669958] [PMC8599732] | ||
| Organizations: |
Leiden University Medical Center, Cell and Chemical Biology, Leiden, South Holland (Netherlands)
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| Purpose: | Tracing cell cycling activities in RPE-fucci cells exposed to various FBS concentrations. | ||||
| Conclusion: | A clear serum concentration-dependent reduction in the proportion of cells arrested in G1 was observed during the cumulative tracing period. | ||||
| Comments: | Cultures of human retinal pigment epithelial cells (RPE-1) with different frequencies of cycling and non-cycling cells were obtained by applying fetal bovine serum (FBS) at various concentrations, i.e., 0, 0.1, 0.5, 1, 5 and 10%. To quantify the frequencies of cells in different cell cycle stages, RPE-1 cells contained the fluorescent ubiquitination-based cell cycle indicator (Fucci) system. In this system, red and green fluorescence reporters fused to the degradation motifs of the cell cycle-controlled Cdt1 and Geminin proteins mark, respectively, cells in G1 and S/G2, respectively. Simultaneous presence of both reporters mark instead cells in early S phase. After seeding and exposing the reporter cells to the various FBS concentrations, cell cycle analysis was performed at the indicated timepoints (12h, 24h and 48h) by flow cytometry. | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Cells cultured in normal condition (10% FBS) served as controls for gating cell populations in G2/M, early S and G1, respectively. At least 10,000 viable singel cells were acquired per sample. | ||||
