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Blood, Vol. 111, Issue 8, 4392-4402, April 15, 2008
A clinical-scale selective allodepletion approach for the treatment of HLA-mismatched and matched donor-recipient pairs using expanded T lymphocytes as antigen-presenting cells and a TH9402-based photodepletion technique
Blood Mielke et al.
111: 4392
Supplemental material for: Mielke et al
Files in this Data Supplement:
- Figure S1. Adaptations of cell and TH9402 concentrations to large-scale conditions (PDF, 90 KB) -
A,B: Variable concentrations of 3-day co-cultured, randomly mismatched responder and irradiated stimulator cells were incubated with variable concentrations of TH9402 for 40 minutes, followed by 90 minutes of extrusion and light exposure of 5 J/cm2 at 180 rpm in the light source. All steps were performed in bags and results were compared to the pre-described26;27 small scale conditions in flasks (1 × 106 viable cells/ml; 10 µM TH9402) as well as an untreated control. Uptake and extrusion of TH9402 was studied in the FITC channel of a flow-cytometer. Analysis of overlay histograms of these experiments showed that increased cell concentrations require lower TH9402 concentrations to reach intracellular TH9402 concentrations after the extrusion phase (directly before light exposure) similar to the small-scale, pre-established condition. C: Overlay histogram of intracellular TH9402 (measured in the FITC channel) for the finally chosen large scale conditions (5 µM and 7.5 µM TH9402 for 5 × 106 viable cells in a bag) in comparison to the pre-established small-scale condition (10 µM TH9402 for 1 × 106 viable cells in a flask). TH: TH9402. MFI: Mean fluorescence intensity.
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