Experiment Overview
| Repository ID: | FR-FCM-Z5L9 | Experiment name: | Rare cells | MIFlowCyt score: | 84.00% |
| Primary researcher: | Jordi Petriz | PI/manager: | Jordi Petriz | Uploaded by: | Jordi Petriz |
| Experiment dates: | 2021-01-07 - 2021-05-30 | Dataset uploaded: | Jul 2022 | Last updated: | Aug 2022 |
| Keywords: | [flow cytometry] [rare cells] [viable DNA-binding dyes] | Manuscripts: |
|
||
| Organizations: |
Germans Trias i Pujol Research Institute, Functional Cytomics, Badalona, Barcelona (Spain)
|
||||
| Purpose: | To present theory and methods to determine the effect of sample manipulation on rare cell detection by validating the Poisson theory and measuring cellular abundance by spiking K562 cells with stable EGFP gene expression at known frequencies. After verifying the applicability of this theory, to evaluate the potential impact of red cell lysis on rare cell detection, either using EGFP-K562 or human leukemia cells. | ||||
| Conclusion: | - Quantification of rare events assessed in terms of cell concentration rather than frequency estimation, may help to achieve robust assays that must be developed and adapted for each patient. - Further experiments will be needed to study the clinical relevance of pathological rare cells in their native sample matrix. - If the malignant rare cell compartment is mainly constituted by ammonium chloride resistant cells, this could lead to an overestimation of CTCs or MRD measurements. - If cell injury derived from sample manipulation can deplete not only healthy, but also pathological cells, including the rare malignant cell compartment, this will also have important implications for the diagnosis and treatment of hematological and nonhematological malignancies. | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Performance Tracking Beads (daily) | ||||
Experiment variables
| Conditions | |
|---|---|
| · K562-EGFP+ | BS7 SAMPLE (PLOT EGFP GATE R7)---2021_099 1% KEG32 1-100 1000 1DCV.fcs · BS7 SAMPLE (PLOT EGFP GATE R7)---2021_099 2% KEG32 1-100 1000 1DCV.fcs · BS7 SAMPLE (PLOT EGFP GATE R7)---2021_099 5% KEG32 1-100 500 1DCV.fcs · BS8 SAMPLE (PLOT EGFP GATE R7)---2021_091 0.1% T2 KEG28 1-100 500 1DCV.fcs · BS9 SAMPLE (PLOT EGFP GATE R7)---2021_096 0.01% T2 KEG29 1-100 1000 1DCV.fcs · 210414 LNW EGFP K562 FILTRAT.fcs · 210414 NLNW EGFP K562 FILTRAT.fcs |
| · K562-EGFP- | BS1 SAMPLE (PLOT EGFP GATE R7)---2021_025 KEG03 2nNEW VF4,5 1-10P x2H DIL 1-100 500.fcs · 2021_229 CLL3 LNW 1% 1-200.fcs · 2021_229 CLL3 NLNW 1% 1-200.fcs · CLL DCV PI CD19-CD5_NLNW VS LNW_2021_229 CLL3 COUNT 1-200.fcs |
| Sample Type | |
|---|---|
| · peripheral blood | BS1 SAMPLE (PLOT EGFP GATE R7)---2021_025 KEG03 2nNEW VF4,5 1-10P x2H DIL 1-100 500.fcs |
| · peripheral blood + K562-EGFP cells | BS7 SAMPLE (PLOT EGFP GATE R7)---2021_099 1% KEG32 1-100 1000 1DCV.fcs · BS7 SAMPLE (PLOT EGFP GATE R7)---2021_099 2% KEG32 1-100 1000 1DCV.fcs · BS7 SAMPLE (PLOT EGFP GATE R7)---2021_099 5% KEG32 1-100 500 1DCV.fcs · BS8 SAMPLE (PLOT EGFP GATE R7)---2021_091 0.1% T2 KEG28 1-100 500 1DCV.fcs · BS9 SAMPLE (PLOT EGFP GATE R7)---2021_096 0.01% T2 KEG29 1-100 1000 1DCV.fcs |
| · K562-EGFP cells | 210414 LNW EGFP K562 FILTRAT.fcs · 210414 NLNW EGFP K562 FILTRAT.fcs |
