Chimerism analysis following nonmyeloablative stem cell transplantation using a new cell subset separation method: Robosep.
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.
Decot
V
V
Cell Therapy Unit and Tissue Bank, CHU Nancy Brabois, Vandoeuvre-Les-Nancy, France.
Latger-Cannard
V
V
Lecompte
T
T
Clément
L
L
Salmon
A
A
Bordigoni
P
P
Stoltz
J-F
JF
Bensoussan
D
D
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*