Successful cryopreservation of purified autologous CD34+ cells: influence of freezing parameters on cell recovery and engraftment

Bone Marrow Transplant. 1998 Dec;22(11):1091-6. doi: 10.1038/sj.bmt.1701494.

Abstract

Conventional hematopoietic stem cell cryopreservation methods use a DMSO concentration of 10%. However, cells manipulated ex vivo may require more refined freezing protocols adapted to the specific cell suspension. In this retrospective study, we evaluated the results obtained with CD34+ cells purified from peripheral blood of 39 patients on the CEPRATE SC System and frozen in 7.5% DMSO with a view to transplantation. The post-freezing recovery of progenitor cells was 89.4 +/- 27.87% for CD34+ cells, 59.13 +/- 36.93% for CFU-GM, and 53.49 +/- 40.71 for BFU-E. Neither the purity of the suspension nor the nucleated cell density during freezing was predictive of cell recovery. No difference was observed between cells stored in vials and bags. Thirty-seven patients transplanted with the concentrated CD34+ fraction received 4.46 x 10(6) CD34+ cells/kg and 33.04 x 10(4) CFU-GM/kg. The median time to granulocyte (>0.5 x 10(9)/l) and platelet (>50 x 10(9)/l) engraftment was 11 and 13 days, respectively. Only cell density and the infused number of CD34+ cells and CFU-GM were significantly related to hematological recovery. Our data suggest that purified CD34+ cells can be successfully cryopreserved in 7.5% DMSO and may represent a first step in establishing freezing parameters for selected CD34+ cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antigens, CD34 / metabolism
  • Cell Survival
  • Colony-Forming Units Assay
  • Cryopreservation / instrumentation
  • Cryopreservation / methods*
  • Cryoprotective Agents
  • Dimethyl Sulfoxide
  • Female
  • Freezing
  • Graft Survival
  • Hematopoiesis
  • Hematopoietic Stem Cell Transplantation*
  • Hematopoietic Stem Cells* / cytology
  • Hematopoietic Stem Cells* / immunology
  • Humans
  • In Vitro Techniques
  • Male
  • Middle Aged
  • Time Factors
  • Transplantation, Autologous

Substances

  • Antigens, CD34
  • Cryoprotective Agents
  • Dimethyl Sulfoxide