Validation of a semi automatic device to standardize quantification of Colony-Forming Unit (CFU) on hematopoietic stem cell products

• Published in: Cytotherapy (Oxford, England) England), 2019-08, Vol.21 (8), p.820-823
• Main Authors: VELIER, MÉLANIE, CHATEAU, ANNE-LINE, MALENFANT, CARINE, OUFFAI, SARAH, CALMELS, BORIS, CHABANNON, CHRISTIAN, LEMARIÉ, CLAUDE
• Format: Article
• Language: English
• Subjects: Advanced Basic Science; Other
• ISSN: 1465-3249; 1477-2566
• DOI: 10.1016/j.jcyt.2019.06.005

AbstractAccurate characterization of hematopoietic stem cells (HSC) products is needed to better anticipate the hematopoietic reconstitution and the outcome in patients. Although CD34+ viable cells enumeration is a key predictor of time to correction of aplasia, it does not fully inform about functionality of cells contained in the graft. CFU assay is the gold standard in vitro potency assay to assess clonogenicity of HSC and consists on the count and identification of colonies several days after culture in a semi solid media. Manual count of colonies with optic microscope is the most commonly used method but its important variability and subjectivity hinders the universal implementation of this potency assay. The aim of this study is to validate a standardized method using the STEMvision™ system, the first semi-automated instrument for imaging and scoring hematopoietic colonies, according to French and European recommendations. Results obtained highlight better performance criteria with STEMvision™ system than the manual method. This semi-automatic device tends to reduce the coefficients of variation of repeatability, inter-operator variability and intermediate precision. This newly available platform could represent an interesting option, significantly improving performances of CFU assays used for the characterization of hematopoietic progenitors.