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.

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

“…Recently, there have been several reports suggesting that cells can be successfully cryopreserved with minimal concentrations of DMSO [39][40][41][42][43][44][45][46][47][48][49][50][51]. For example, Zhao et al [46] showed that the use of 5% or 10% DMSO in combination with either 20% or 70% serum produced similar results during cryopreservation of fetal human liver CD34 + cells.…”

Clinical grade adult stem cell banking

“…A review of the published literature over the past several years identifies several reports suggesting that adult stem cells (hematopoietic and non-hematopoietic) can be successfully cryopreserved with minimal concentrations of DMSO. 45,[52][53][54][55][56][57][58][59][60][61][62] For example, Zhao et al 57 showed that the use of 5 or 10% DMSO produced similar results during cryopreservation of fetal human liver CD34 + cells. Sputtek et al 60 reported optimal cryopreservation of peripheral blood HPCs using at least 5% DMSO.…”

Variation in dimethyl sulfoxide use in stem cell transplantation: a survey of EBMT centres

“…It has been suggested that lowering the dose of DMSO may be advisable. Lower concentrations, for example 5% DMSO 16,17 or 3.5% DMSO 18 are an effective cryoprotectant. Stem cell washing to remove DMSO appears to have no adverse effect on haematological recovery and may reduce toxicity 19 and fractionation of stem cell administration may similarly reduce side effects.…”