ABSTRACT
BACKGROUND: JACIE Standards (FACT Standards in the USA) have been implemented in Europe since 1999. An on-site accreditation inspection took place at our center in January 2004. The purpose of this work was to develop a real-time process/quality control system meeting the JACIE Standards for HPC release. METHODS: Data from 194 HPC processing procedures for autologous transplantation performed over a 5-year period were analyzed. The results of different processing methods applied at our facility were compared: (1) cryopreservation without washing cells (n=50), (2) washing cells (n=87), (3) cell-density separation (n=12) and (4) positive CD34 selection (n=45). RESULTS: Four critical control points were set for the validation of HPC processing: (a) number of lost CD34(+) cells during processing, (b) contamination, (c) viability of the cells after thawing and (d) ability to reconstitute hematopoiesis after transplantation. On the basis of statistical analysis, ranges of acceptable values were defined for each critical control point and for each processing method. Those acceptable values were used for cell release and real-time quality control. DISCUSSION: This study describes a model for the validation of HPC processing and for a real-time process/quality control system for HPC release. Optimization of processing techniques, standardization of methods and comparison between facilities will open the way towards external quality controls and quality improvement.
Subject(s)
Hematopoietic Stem Cell Transplantation/standards , Antigens, CD34/analysis , Cryopreservation , Humans , Quality Control , Time Factors , Transplantation, AutologousSubject(s)
Antigens, CD34/metabolism , Blood Cells/metabolism , Blood Component Removal/instrumentation , Blood Component Removal/methods , Blood Specimen Collection/methods , Adolescent , Blood Cells/cytology , Blood Component Removal/standards , Cell Separation/instrumentation , Cell Separation/methods , Cell Separation/standards , Child , Child, Preschool , Efficiency , Humans , InfantABSTRACT
In children, only a few guidelines are available for optimizing peripheral blood progenitor cell (PBPC) harvesting. We analyzed by means of flow cytometry and clonogenic assays 60 harvest products obtained from 20 children by standardized leukapheresis after treatment with chemotherapy and CSF. In addition, 27 fresh blood samples obtained prospectively during the mobilization phase were studied. CFU-GM/kg significantly correlated with MNC/kg, CD34+ cells/kg and CD34+33- cells/kg in apheresis products (P < 0.001). In fresh blood samples, CFU-GM/ml significantly correlated with MNC/ml, CD34+ cells/ml and CD34+33- cells/ml (P < 0.001). The numbers of CD34+ cells/ml, CD34+33- cells/ml and MNC/ml in 19 blood samples taken prior to leukapheresis were compared with CFU-GM/kg harvested and thawed after cryopreservation applying multiple regression analysis with stepwise variable selection. The number of circulating CD34+ cells/ml prior to leukapheresis highly correlated with and was predictive for the number of collected CFU-GM/kg (P < 0.001). In addition, a significant correlation (P < 0.05) between the number of progenitor cells/kg reinfused and the time to myeloid and platelet recovery was found in children undergoing high-dose therapy. Our data indicate that a single leukapheresis will be sufficient to obtain a minimum number of 5 x 10(4) CFU-GM/kg if the pre-harvest number of circulating CD34+ cells is > or = 10(5)/ml. Thus, our results will help to optimize PBPC transplantation in children.
