[Erythrocyte removal from bone marrow by density gradient separation using the COBE 2991 cell processor with the triple-bag processing set].
ABO-incompatible bone marrow transplantation requires red blood cell depletion. Lots of laboratory adopted the technique of density gradient centrifugation (Ficoll-hypaque) using the COBE 2991 cell processor with simple-bag processing set. However, tubing of this set is not adapted to the currently available peristaltic pumps. Moreover, two other sets are required: one for the buffy-coat and one for postgradient cell washing. We developed a method using triple-bag processing set to conduct whole-step procedure (concentration, Ficoll and washing). Peristaltic PVC tubing is provided in one line of the set allowing a safe processing without several connections thus reducing risks of microbial contamination. First, we used buffy-coat of total blood for training, then, we carried out red cell depletion of healthy bone marrow donors. The red blood cell depletion was 97.9+/-1.1% and CD34+ recovery was 89.6+/-8.7%. These results are very close to those obtained with the simple-bag set (red cell depletion.=94.0+/-6.8% and CD34+ recovery=95.9+/-20.3%). We conclude that the triple-bag system, very little used in France, is practical, simplified the manipulation and is more safety than the simple-bag set.
Cappellesso-Fleury
S
S
Laboratoire de thérapie cellulaire, établissement français du sang Pyrénées-Méditerranée, 31300 Toulouse, France. sandrine.fleury@efs.sante.fr
Rage
C
C
Tschaggeny
F
F
Gaudé
J
J
Gomez
M
M
Bourin
P
P
D000017:ABO Blood-Group System; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D004912:Erythrocytes; D006801:Humans; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000295:instrumentation* / Q000379:methods
D000017:ABO Blood-Group System