title: A new system for quality control in hematopoietic progenitor cell units before reinfusion in autologous transplant.
abstract: BACKGROUND
  In our Center, the cell viability, the integrity of the bag, and the clonogenic assay were evaluated before the reinfusion of hematopoietic progenitor cells-apheresis (HPC-A). This quality control (QC) should be made 14 days before the reinfusion to the patient to have the result of the functional test on the proliferative capacity of hematopoietic progenitors.
  
  
  METHODS
  This study was designed to assess the potential of an automatic cell counting system (NucleoCounter NC-3000, ChemoMetec) in our clinical routine as a support of the clonogenic assay and the cytofluorimetric analysis for the QC of the cryopreserved HPC-A. The cell viability was evaluated by flow cytometry using the modified International Society of Hematotherapy and Graft Engineering protocol. The proliferative potential was assessed by specific clonogenic tests using a commercial medium. Furthermore, we evaluated the cellular functionality with NucleoCounter NC-3000, by using two protocols: "vitality assay" and "mitochondrial potential assay."
  
  
  RESULTS
  The evaluation of the total nucleated cells in preapoptosis measured by 5,5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazol-carbocyanine iodide (JC-1) assay showed a negative correlation (r=-0.43) with the total number of colonies (colony-forming unit [CFU]-granulocyte-macrophage progenitors plus burst-forming unit-erythroid progenitors plus CFU-granulocyte, erythroid, macrophage, megakaryocyte progenitors) obtained after seeding of 50 × 10(6) /L viable total nucleated cells. We observed a significant difference (p<0.0001) comparing the median number of colonies (166.70; SD, ± 136.36) obtained with a value of JC-1 less than 30% to the number of colonies (61.75; SD, ± 59.76) obtained with a value of JC-1 more than 30%.
  
  
  CONCLUSIONS
  The evaluation of cell functionality by the use of the NucleoCounter NC-3000 is in agreement with results from clonogenic assay and can be considered an effective alternative in the routine laboratory.
authors.affiliation: Cryolab Center of Biotechnology and Cryobiology, Immunohematology Section, SIMT, Department of Hematology, Tor Vergata University, Rome, Italy; Rome Transplant Network, Department of Hematology, Tor Vergata University, Rome, Italy.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Scerpa
authors.forename: Cecilia
authors.initials: C
authors.lastname: Rossi
authors.forename: Nicola
authors.initials: N
authors.lastname: Daniele
authors.forename: Alessandro
authors.initials: A
authors.lastname: Lanti
authors.forename: Gaspare
authors.initials: G
authors.lastname: Adorno
authors.forename: Alessandra
authors.initials: A
authors.lastname: Picardi
authors.forename: William
authors.initials: W
authors.lastname: Arcese
authors.forename: Sergio
authors.initials: S
authors.lastname: Amadori
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Isacchi
authors.forename: Francesco
authors.initials: F
authors.lastname: Zinno
mesh_terms: D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D015925:Cryopreservation; D005434:Flow Cytometry; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D011786:Quality Control; D014182:Transplantation, Autologous / Q000379:methods*