Cryopreservation of HPCs with high cell concentration in 5‐percent DMSO for transplantation to children

Abstract

Transfusion of cryopreserved HPCs carries a risk of serious adverse effects due to the use of DMSO. Some studies have shown that the incidence and severity of adverse reactions are dose related. 1 It is desirable therefore to reduce the amount of DMSO in cryopreserved components to minimize toxicity, particularly in children.The concentration of DMSO and the concentration of cells to be cryopreserved determine the volume and the amount of DMSO infused. Previous studies 2,3 describe satisfactory results using 5-percent DMSO or 3.5percent DMSO with HES. Also, high cell concentrations (up to 800 ‫ן‬ 10 6 /mL) in 10-percent DMSO have not shown a deleterious impact on function or engraftment of HPCs, either in vitro or in vivo. 4 Our approach used a high cell concentration (>200 ‫ן‬ 10 6 /mL) and a low DMSO concentration (5 percent) for cryopreservation. We report hematologic recovery in patients who received such transplants.HPCs were collected from 13 consecutive patients for autologous transplant. The cells were cryopreserved with 5-percent DMSO in autologous plasma as the only cryoprotectant at a final cell concentration equal to or greater than 200 ‫ן‬ 10 6 per mL. The cells were frozen at -80ЊC by immersion in a methanol bath in a mechanical freezer, 5 and stored at that same temperature. All patients were conditioned with myeloablative regimens and medicated with corticosteroids, antihistamines, and diuretics prior to transfusion of their thawed, unmanipulated HPCs.There were 6 boys and 7 girls with a median age of 11.8 years (range 4.5-19.2). Diagnoses were brain tumor in 5, Ewing sarcoma in 3, Hodgkin disease in 2, neuroblastoma in 1, T-cell acute lymphoblastic leukemia in 1, and Burkitt lymphoma in 1.The median volume of the cryopreserved HPCs and their cell concentrations were 300 mL (range, 134-700) and 263 ‫ן‬ 10 6 per mL (range, 192-390), respectively, with a median CD34+ cell content of 3.66 ‫ן‬ 10 6 per kg (range, 2.00-6.44). Median time from cryopreservation to transplantation was 10 days (range, 9-108).The median dose of DMSO infused was 0.41 mL per kg (range, 0.13-0.58) in an HPC component with a median volume of 8.2 mL per kg (range, 2.7-11.7). Adverse effects during transfusion were minimal. One patient experienced abdominal pain and vomiting and another required treatment for hypertension with nifidipine. If the HPCs had been cryopreserved by standard protocols (cell concentration 100 ‫ן‬ 10 6/ mL in 10 percent DMSO), the median volume of DMSO and HPC component transfused would have been 1.8 mL per kg (range 0.70-4.55) and 18 mL per kg (range 7-45), respectively.A granulocyte count > 0.5 ‫ן‬ 10 9 per L and a platelet count > 20 ‫ן‬ 10 9 per L were achieved after a median of 11 (range 10-14) and 12 (range 10-23) days, respectively. Hematologic recovery was adequate (Hb > 100 g/L, platelets > 100 ‫ן‬ 10 9/ L, and granulocytes > 1 ‫ן‬ 10 9/ L) in 9 of 12 evaluable patients at 3 months and in 9 of 10 evaluable patients at 6 months post-transplant. The only patient not achieving the target values was ...

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.…”

Autologous Stem Cell Transplantation in the Treatment of Amyloidosis – Can Manipulation of the Autograft Reduce Treatment-Related Toxicity?

“…Using DMSO concentrations lower than 10 percent reduces complication rate (Akkok et al, 2009;Curcoy et al, 2002;Galmes et al, 2007;Windrum et al, 2005). There are only a few clinical studies where DMSO is used as the sole cryoprotectant at a 5 percent or lesser concentration (Akkok et al, 2009;Curcoy et al, 2002;Galmes et al 1996Galmes et al , 2007.…”

“…There are only a few clinical studies where DMSO is used as the sole cryoprotectant at a 5 percent or lesser concentration (Akkok et al, 2009;Curcoy et al, 2002;Galmes et al 1996Galmes et al , 2007. One other group used 3.5% DMSO, which is the lowest reported DMSO concentration in a clinical study but it was used in combination with 2.5% hydroxyethyl starch and followed by storage at -80°C (Halle et al, 2001).…”

“…However, the authors do not recommend storage of grafts at -80°C for longer than 6 months due to progressively diminishing viability of mononuclear cells (MNC) and recovery rates of colony-forming units; granulocyte-macrophage (CFU-GM) and burst-forming unit erythroid (BFU-E) reaching zero after 24 months, when grafts are cryopreserved with 5% DMSO (Galmes et al, 2007). In a pediatric setting, Curcoy and colleagues reported successful engraftment after short time storage at -80°C by using only 5% DMSO (Curcoy et al, 2002). It was also illustrated by in vitro studies that the numbers of both total and viable CD34+ cells were higher, CD34+ cells were less apoptotic and necrotic plus early hematopoietic progenitor cells were better preserved with 5 compared to 10% DMSO (Abrahamsen et al, 2002).…”

Are orange lollies effective in preventing nausea and vomiting related to dimethyl sulfoxide? A multicenter randomized trial

“…The toxicity is proportional to the concentration and amount of DMSO [7] and increases with the number of bags and of cells harvested and with the patient's weight [2,4,6,8]. A 10% solution is generally used [2,3]; the 5% solution with lower incidence of side effects is not recommended for lengthy storage since its safety has been assessed only over a few months [9,10].…”