title: Chimerism analysis following nonmyeloablative stem cell transplantation using a new cell subset separation method: Robosep.
abstract: Chimerism analysis has become an important tool to manage patients in the peri-transplant period of allogenic stem cell transplantation. During this period, cells of donor and host origin can coexist and increasing proportion of cells of host origin is considered as a recurrence of the underlying disease. We currently performed chimerism analysis on separate peripheral blood cell subsets, lymphocytes and granulocytes. To improve our isolation method, a new automated device from Stem Cell Technology Roboseptrade mark was tested and compared to our manual separation technique. The results obtained on T cell purification showed an improvement of the purity (98.42% with Robosep vs. 92.42% with the manual technique Rosettesep) and of the recovery (63.43% with Robosep and 38% with Rosettesep). The results were significantly improved on patient samples with less than 10% CD3 positive cells (purity: 90% vs. 44.44%; recovery: 73.79% vs. 43.98%). Granulocytes separation was based on CD15 expression. The results showed an improvement of the purity with Robosep (96.90% vs. 86.20% with the manual technique Polymorphprep) but the recovery was impaired (35.2% vs. 52.30%). Using a myeloid (CD66/CD33) cocktail, recovery was improved with the Robosep device (64.04% with the myeloid cocktail vs. 22.4% with the CD15 cocktail). Our data demonstrated that Robosep allowed a performant cell purification in the early period post-transplantation even for populations representing less than 10% of the peripheral blood cells.
authors.affiliation: Cell Therapy Unit and Tissue Bank, CHU Nancy Brabois, Vandoeuvre-Les-Nancy, France.
authors.forename: V
authors.initials: V
authors.lastname: Decot
authors.forename: V
authors.initials: V
authors.lastname: Latger-Cannard
authors.forename: T
authors.initials: T
authors.lastname: Lecompte
authors.forename: L
authors.initials: L
authors.lastname: Clément
authors.forename: A
authors.initials: A
authors.lastname: Salmon
authors.forename: P
authors.initials: P
authors.lastname: Bordigoni
authors.forename: J-F
authors.initials: JF
authors.lastname: Stoltz
authors.forename: D
authors.initials: D
authors.lastname: Bensoussan
mesh_terms: D001772:Blood Cell Count / Q000379:methods*; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D006098:Granulocytes / Q000473:pathology*; D006801:Humans; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000473:pathology*; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000473:pathology*