title: A simplified method for cryopreservation of peripheral blood stem cells at -80 degrees C without rate-controlled freezing.
abstract: A simplified method was established for cryopreservation of peripheral blood stem cells (PBSCs) using hydroxyethyl starch (HES) and dimethylsulfoxide (DMSO) as a cryoprotective agent at -80 degrees C without rate-controlled freezing. The data indicate that a cryoprotective solution consisting of 6% HES and 5% DMSO produced the highest recovery rates for nucleated cells (92.0 +/- 3.5%), CFU-GM (73.8 +/- 4.1%) and BFU-E (82.2 +/- 6.9%), and the highest trypan blue viability (88.4 +/- 3.6%). For long-term cryopreservation of PBSCs, CFU-GM recovery rates remained almost unchanged during 5-18 months; the mean CFU-GM recovery rate after 18 months of cryopreservation was 70 +/- 11%. When large-scale samples of PBSCs in 100-ml freezing bags were cryopreserved for clinical use, cell and CFU-GM recoveries were similar. Using this method, 10 patients with hematological malignancy received PBSC transplants after marrow-ablative chemotherapy. All demonstrated early engraftment; seven are now alive in unmaintained complete remission for 3.5-15 months after PBSC transplant. This simple and inexpensive method will be useful for the wider application of PBSC transplant as a therapeutic alternative in the treatment of malignant diseases.
authors.affiliation: Blood Transfusion Service, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Makino
authors.forename: M
authors.initials: M
authors.lastname: Harada
authors.forename: K
authors.initials: K
authors.lastname: Akashi
authors.forename: S
authors.initials: S
authors.lastname: Taniguchi
authors.forename: T
authors.initials: T
authors.lastname: Shibuya
authors.forename: S
authors.initials: S
authors.lastname: Inaba
authors.forename: Y
authors.initials: Y
authors.lastname: Niho
mesh_terms: D000208:Acute Disease; D000328:Adult; D016026:Bone Marrow Transplantation; D002675:Child, Preschool; D003114:Colony-Forming Units Assay; D015925:Cryopreservation / Q000379:methods*; D004121:Dimethyl Sulfoxide; D005069:Evaluation Studies as Topic; D005260:Female; D006412:Hematopoietic Stem Cells; D006801:Humans; D006895:Hydroxyethyl Starch Derivatives; D007938:Leukemia / Q000601:surgery; D007963:Leukocytes, Mononuclear; D008297:Male; D009447:Neuroblastoma / Q000601:surgery
chemical_list: D006895:Hydroxyethyl Starch Derivatives; D004121:Dimethyl Sulfoxide