Impact of uncontrolled freezing and long-term storage of peripheral blood stem cells at −80 °C on haematopoietic recovery after autologous transplantation. Report from two centres

Abstract

Controlled-rate freezing and storage in vapour phase nitrogen are used by most transplantation teams for the cryopreservation and storage of peripheral blood haematopoietic stem cells (PBSC). In this study, we analysed 666 autologous PBSC transplants after uncontrolled freezing and storage of PBSC at − 80°C. Statistical analysis showed that neutrophil recovery was associated with both the infused CD34 + cell dose (P = 0.01) and the post transplantation use of growth factors (P o 0.001) and that platelet recovery was associated with the infused CD34 + cell dose (P o0.001) and with the diagnosis (P = 0.02). We analysed three groups according to the duration of the cryopreservation period (less than 6 months, between 6 and 12 months or more than 1 year). Haematopoietic recovery was not found to be adversely affected by longer storage at − 80°C. The haematopoietic recoveries of 50 pairs of sequential transplantations from the same PBSC mobilization were analysed. Despite prolonged cryopreservation, there were no statistically significant differences in neutrophil (P = 0.09) or platelet (P = 0.22) recovery in the second compared with the first transplant. In conclusion, the long-term storage of PBSC at − 80°C after uncontrolled-rate freezing is an easy and comparatively inexpensive cryopreservation method that leads to successful haematopoietic recovery even after prolonged storage.

Cryopreservation of Hematopoietic Stem Cells: Emerging Assays, Cryoprotectant Agents, and Technology to Improve Outcomes

“…Sputtek et al [73] compared the recovery of PBSCs frozen at –80°C versus –170°C for 3 and 6 months and found that –80°C storage resulted in significant loss of cell membrane integrity and clonogenic potential. Encouraging results from one study found that hematopoietic recovery for PBSCs was found not adversely affected by long storage (> 1 year) at –80°C, which was demonstrated at two independent centers using similar freezing and storage techniques [74]. Yuan et al [75] found success at stabilizing pluripotent stem cells for up to 1 year at –80°C by adding the minimally toxic polysaccharide Ficoll 70 to their cryoprotectant solution.…”

Supportive (Maintenance) Therapy in Hematopoietic Stem Cell Transplantation: Main Principles and New Perspectives

“…В исследовании G. Detry и соавт. применение упрощенного метода неконтролируемого замораживания и хранения периферических ГСК при температуре -80 °C (n = 666) с использованием 5% ДМСО совместно с гидроксиэтилкрахмалом в качестве внеклеточного криопротектора также способство-вало успешному восстановлению гемопоэза после длительного (> 12 мес., n = 60) хранения [42].…”

Preserved in vivo reconstitution ability of PBSCs cryopreserved for a decade at −80 °C

“…Other studies also showed rapid and sustained engraftment after autologous PBSCT stored at − 80°C with the simplified method for several months to years. 18,19,21 McCullough et al 7 compared the impacts of LT storage of PBSCs up to 5 years between the two cryopreservation methods; viability and recovery of CD34 + cells were comparable. Our study confirms and extends these studies by evaluating recovery rates of CD34 + cells and their functions including differentiation, reconstitution and self-renewal after more than 10 years cryopreservation at − 80°C.…”

“…[15][16][17] Regarding uncontrolled freezing and storage at − 80°C, several studies have shown that ST storage for months to several years does not decrease viability of CD34 + cells or colony-forming cells. 5,7,[18][19][20] However, few studies have addressed effects of LT storage of PBSC products. Costa et al 17 reported that viability of TNCs was around 80% and not changed with storage time up to 14 years.…”

“…7,[17][18][19][20] The mechanical freezing method at − 80°C provides technical simplification and cost reduction, and therefore supports a hematopoietic stem cell transplant program in countries with limited resources in startup conditions as an alternative to liquid nitrogen storage, but requires a constant and reliable electrical power supply. However, our results need to be confirmed by clinical engraftment data or in future prospective studies to compare viability and recovery between bags cryopreserved at − 80°C or in liquid nitrogen from the same donors.…”