title: Successful cryopreservation of purified autologous CD34+ cells: influence of freezing parameters on cell recovery and engraftment.
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.
authors.affiliation: Laboratoire de Thérapie Cellulaire, Centre de Transfusion Sud Est Francilien, Hôpital Henri Mondor, Créteil, France.
authors.forename: F
authors.initials: F
authors.lastname: Beaujean
authors.forename: J H
authors.initials: JH
authors.lastname: Bourhis
authors.forename: C
authors.initials: C
authors.lastname: Bayle
authors.forename: H
authors.initials: H
authors.lastname: Jouault
authors.forename: M
authors.initials: M
authors.lastname: Divine
authors.forename: C
authors.initials: C
authors.lastname: Rieux
authors.forename: M
authors.initials: M
authors.lastname: Janvier
authors.forename: C
authors.initials: C
authors.lastname: Le Forestier
authors.forename: J L
authors.initials: JL
authors.lastname: Pico
mesh_terms: D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism; D002470:Cell Survival; D003114:Colony-Forming Units Assay; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D005260:Female; D005615:Freezing; D006085:Graft Survival; D006410:Hematopoiesis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D013997:Time Factors; D014182:Transplantation, Autologous
chemical_list: D018952:Antigens, CD34; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide