title: Density-gradient separation for 4-hydroperoxycyclophosphamide purging of autologous bone marrow grafts.
abstract: We studied density-gradient separation of autologous bone marrow grafts in preparation for ex vivo 4-hydroperoxy-cyclophosphamide (4-HC) purging. A two-step procedure of buffy coat isolation followed by Ficoll-diatrizoate separation was used. Buffy-coat cells were isolated primarily either using the COBE 2991 Blood Cell Processor or Haemonetics 30 Cell Separator. All density-gradient separations were performed using the COBE 2991. The nucleated cell recoveries after buffy-coat isolation were 44.3 +/- 10.4% and 85.1 +/- 10.5% (+/- standard deviation, P less than 0.001) for the Haemonetics 30 and COBE 2991 isolated grafts, respectively. The final nucleated cell recoveries after density-gradient separation for these two buffy-coat techniques were 24.2 +/- 10.1% and 29.5 +/- 14.7% (p = 0.30). Compared to manual density-gradient separation, the techniques using the COBE 2991 recovered slightly higher numbers of nucleated cells and CFU-GM. The red blood cell concentration of the 4-HC incubation mixture was 1.0 +/- 0.6% and the CFU-GM survival after treatment was 2.4 +/- 2.6%. The variability in CFU-GM survival after 4-HC incubation of the density-gradient separated grafts appears less than previous experience treating buffy-coat cells. Density-gradient separation of autologous bone marrow appears to provide a more uniform treatment with 4-HC and may improve the clinical transplant course of the autograft recipient.
authors.affiliation: Johns Hopkins Oncology Center, Baltimore, Maryland 21205.
authors.forename: S
authors.initials: S
authors.lastname: Rowley
authors.forename: J
authors.initials: J
authors.lastname: Davis
authors.forename: H
authors.initials: H
authors.lastname: Braine
authors.forename: R
authors.initials: R
authors.lastname: Jones
authors.forename: A
authors.initials: A
authors.lastname: Yeager
authors.forename: R
authors.initials: R
authors.lastname: Saral
authors.forename: G
authors.initials: G
authors.lastname: Santos
mesh_terms: D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D015925:Cryopreservation; D003520:Cyclophosphamide / Q000031:analogs & derivatives* / Q000627:therapeutic use; D004906:Erythrocyte Count; D006801:Humans; D014182:Transplantation, Autologous
chemical_list: D003520:Cyclophosphamide; C011272:perfosfamide