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.


OBJECTIVE
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
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.


CONCLUSIONS
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.
SIMT, IRCCS Bambino Gesù Pediatric Hospital, Rome, Italy. mcscerpa@yahoo.it
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Locatelli
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D018952:Antigens, CD34; D017209:Apoptosis; D001331:Automation; D002452:Cell Count / Q000379:methods; D003257:Consumer Product Safety; D015925:Cryopreservation / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods* / Q000592:standards; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D009336:Necrosis; D015203:Reproducibility of Results
D018952:Antigens, CD34