For co-culture experiments with Luciferase-expressing B-ALL PDX samples, 3x104/cm2 mesenchymal stem cells (MSC) were plated in 96-well tissue culture plates and 2x104 B-ALL PDX cells were added in StemSpan Serum-Free Expansion Medium II (SFEM II, STEMCELL Technologies, Cambridge, UK).17 (link),18 (link) Cells were treated for 5 days with indicated drugs and luminescence analyzed with the Steady-Glo Luciferase Assay System (Promega, Southampton, UK), according to manufacturer’s instruction, and detected with Infinite m200 Pro microplate reader (Tecan, Reading, UK). The following reagents and inhibitors were used, S3I-201 (Cayman Chemical, MI, USA), Napabucasin (BBI608; MedChemExpress, NJ, USA), C188-9 (MedChemExpress and Adooq Biosciences, CA, USA), Nutlin-3a (Merck Life Science, Dorset, UK) and Idasanutlin (Bio-Techne, Abingdon, UK).
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Co-culture of B-ALL PDX with MSCs
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Corresponding Organization :
Other organizations : University College London, Great Ormond Street Hospital, Royal Marsden NHS Foundation Trust, Northumbria University, Princess Máxima Center, Radboud University Nijmegen, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center
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Variable analysis
independent variables
- S3I-201
- Napabucasin
- C188-9
- Nutlin-3a
- Idasanutlin
- Luminescence
- 3x10^4/cm^2 mesenchymal stem cells (MSC) plated in 96-well tissue culture plates
- 2x10^4 B-ALL PDX cells added in StemSpan Serum-Free Expansion Medium II (SFEM II)
- Cells treated for 5 days with indicated drugs
- Positive control: Not specified
- Negative control: Not specified
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