Automated washing of human progenitor cells: evaluation of apoptosis and cell necrosis

Background: High-dose chemotherapy followed by reinfusion of autologous stem cells harvested from peripheral blood has been increasingly applied for a variety of disorders. The critical importance of cell dose in the clinical outcome, after transplant, has motivated the need to develop techniques aimed at reducing cell losses and increasing reproducibility.Objectives: The aim of this study is to evaluate the efficacy of the Sepax S-100 device to process thawed HPC-A products in comparison with manual procedure.Methods/Materials: We have analysed viability, total nucleated cells (TNC), haematopoietic progenitors and CD34+ cells recovery.Results: The TNC and CD34+ cells recovery in the automatic procedure was of 91.9% (73-100; SD +/- 12.60) and 86.7% (69-100; SD +/- 10.21), respectively. Instead the recovery of TNC and CD34+ cells using the manual method was of 84.7% (47-100; SD +/- 22.9) and 80.29% (23-100; SD +/- 25.96). The results, obtained from the assessment of viability of CD34+ both 7-AAD)+ and AnnV+ showed a high percentage of necrosis and apoptosis in this cell subset by using the manual procedure in respect to the Sepax automated system.Conclusion: Overall, our data suggest that the automated washing procedure is safe and suitable for processing of thawed HPC-A products and can be daily used in clinical routine.