title: No differences in colony formation of peripheral blood stem cells frozen with 5% or 10% dimethyl sulfoxide.
abstract: High-dose chemotherapy with autologous stem cell rescue usually requires cryopreservation of the cells. For several years, 10% dimethyl sulfoxide (DMSO) has been used as the standard cryoprotectant. Because DMSO infusion can lead to toxic clinical complications in a dose-related manner, we wanted to evaluate if reduction to 5% DMSO would be possible. We have compared colony formation in the myeloid, erythropoietic, and megakaryocyte lineages in peripheral blood progenitor cell (PBPC) samples cryopreserved in parallel with 5% and 10% DMSO. Twenty-seven PBPC samples from patients with malignant diseases were investigated after 3 months of cryopreservation in liquid N(2), and samples from 14 of these patients were investigated after 1 year. A significantly higher colony formation was demonstrated for colony-forming units-erythrocyte (CFU-E) and CFU-granulocyte, erythrocyte, macrophage, megakaryocyte (GEMM) both at 3 months and at 1 year in the 5% samples. For CFU-granulocyte-macrophage (GM) and CFU-megakaryocyte (Mk) no significant difference was demonstrated neither at 3 months nor at 1 year in samples frozen with 5% and 10% DMSO. Also, there was a statistically significant correlation between the CFU-total and CFU-Mk-total, indicating that the CFU-total might be used as an evaluation of megakaryocyte progenitors. Viability testing with the Trypan Blue exclusion test showed that cells cryopreserved in 5% DMSO had significantly higher viability than the cells cryopreserved in 10% DMSO. We conclude that 5% DMSO is at least as good for cryopreservation of small-volume PBPC samples as the conventional 10% DMSO, and our results suggest that the possibility of using 5% DMSO for cryopreservation of autologous PBPC grafts should be further investigated in clinical studies.
authors.affiliation: The Blood Bank, Haukeland University Hospital, Bergen, Norway. anne.bakken@helse-bergen.no
authors.forename: A M
authors.initials: AM
authors.lastname: Bakken
authors.forename: O
authors.initials: O
authors.lastname: Bruserud
authors.forename: J F
authors.initials: JF
authors.lastname: Abrahamsen
mesh_terms: D001859:Bone Neoplasms / Q000097:blood; D002455:Cell Division / Q000187:drug effects*; D003114:Colony-Forming Units Assay; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D006689:Hodgkin Disease / Q000097:blood; D006801:Humans; D008228:Lymphoma, Non-Hodgkin / Q000097:blood; D008297:Male; D009101:Multiple Myeloma / Q000097:blood; D012509:Sarcoma / Q000097:blood; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D013736:Testicular Neoplasms / Q000097:blood; D014182:Transplantation, Autologous
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide