Autologous bone marrow transplantation using unfractionated cells cryopreserved in dimethylsulfoxide and hydroxyethyl starch without controlled-rate freezing

Abstract

To develop a simplified method of bone marrow (BM) cryopreservation, changes were made in the standard method in three areas: the cryoprotectant, the method of cell freezing, and the storage temperature. Unfractionated BM cells from 60 patients were cryopreserved in 300-mL aliquots in both dimethylsulfoxide (DMSO) and hydroxyethyl starch (HES), a combination known to preserve granulocyte successfully. The cells were frozen without rate-controlled freezing by simple immersion into a -80°C freezer where they remained until the time of reinfusion. The 60 patients underwent 72 autologous transplants after three high-dose chemotherapy regimens: 30 received high-dose carmustine in combination, five received high-dose busulfan and cyclophosphamide, and 37 received high-dose aziridimylbenzoquinone. The BM was infused for more than 30 minutes after rapid thawing at 37°C. The mean post-thaw nucleated cell recovery was 96% ± 11.6%, and Trypan blue dye exclusion was 82.2% ± 9.2%. The mean postthaw CFU-GM and BFU-E recoveries were 81.9% ± 39.0% and 90.5% ± 41.2%. Complete count recovery occurred in 68 of 72 transplants. Median times to a WBC count > 1,000/μL, a granulocyte count > 1,000/μL and a platelet count > 20,000/μL were 15, 21, and 15 days, respectively. Risk factors for delayed recovery were not found. Unfractionated BM cells can be successfully cryopreserved in the DMSO/HES mixture rapidly and inexpensively, without rate-controlled freezing or storage at liquid nitrogen temperatures.