Table 2.
Semiclosed, clinical-scale photodepletion and automated Ficoll (N = 3)
| Exp no. | Stim put in coculture TNCs, ×109 | Resp put in coculture TNCs, ×109 | TNCs in coculture at start, ×109 | TNCs in coculture after 72 h, ×109 | TVCs in coculture after 72 h, ×109 | Research sample TVCs, ×109 | Cells proceeded to PD TVCs, ×109 | Final SD product after wash and Ficoll TNCs, ×109 | Viability, % | CD3+ cells, % | Total viable CD3+ cells, ×109 | SD transplantation dose yield for a 70-kg patient, ×107/kg |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| VII | 5.40 | 5.40 | 10.80 | 7.80 | 5.40 | 0.40 | 5.00 | 1.82 | 91 | 72 | 1.19 | 1.70 |
| VIII | 12.00 | 12.00 | 24.00 | 17.30 | 9.90 | 3.90 | 6.00 | 1.43 | 87 | 81 | 1.01 | 1.44 |
| IX | 9.20 | 9.20 | 18.40 | 15.80 | 9.80 | 3.80 | 6.00 | 0.98 | 85 | 87 | 0.73 | 1.04 |
| Median | 9.20 | 9.20 | 18.40 | 15.80 | 9.80 | — | 6.00 | 1.43 | 87 | 81 | 1.01 | 1.44 |
With regard to clinical feasibility and to improve viability of final selectively depleted products, photodepleted cells were washed and separated using a semiclosed, automated Ficoll Hypaque density gradient separation step directly after PD (day +0, Exp VII-IX). PD was performed in a single session using 2 bags (for 5.0 μM and 7.5 μM TH9402) with a maximum cell content of 3 × 109 viable cells per bag. For freshly obtained cell products, total nuclear cell counts were obtained. For cultured or treated products, total viable cell counts were obtained.
Stim indicates irradiated stimulator cell (expanded T lymphocytes); TNCs, total nucleated cells; Resp, responder cell; TVCs, total viable cells; and —, not available.