Beaujean F, Bourhis J H, Bayle C, Jouault H, Divine M, Rieux C, Janvier M, Le Forestier C, Pico J L
Laboratoire de Thérapie Cellulaire, Centre de Transfusion Sud Est Francilien, Hôpital Henri Mondor, Créteil, France.
Bone Marrow Transplant. 1998 Dec;22(11):1091-6. doi: 10.1038/sj.bmt.1701494.
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.
传统的造血干细胞冷冻保存方法使用的二甲基亚砜（DMSO）浓度为10%。然而，体外操作的细胞可能需要更精细的冷冻方案，以适应特定的细胞悬液。在这项回顾性研究中，我们评估了从39例患者外周血中纯化得到的CD34+细胞，在CEPRATE SC系统中用7.5% DMSO冷冻以备移植的结果。冷冻后祖细胞的回收率对于CD34+细胞为89.4±27.87%，对于粒-巨噬细胞集落形成单位（CFU-GM）为59.13±36.93%，对于爆式红系集落形成单位（BFU-E）为53.49±40.71。冷冻过程中悬液的纯度和成核细胞密度均不能预测细胞回收率。在小瓶和袋子中储存的细胞之间未观察到差异。37例接受浓缩CD34+组分移植的患者，每千克体重接受4.46×10⁶个CD34+细胞和33.04×10⁴个CFU-GM。粒细胞（>0.5×10⁹/L）和血小板（>50×10⁹/L）植入的中位时间分别为11天和13天。只有细胞密度以及输注的CD34+细胞和CFU-GM数量与血液学恢复显著相关。我们的数据表明，纯化的CD34+细胞可以成功地在7.5% DMSO中冷冻保存，这可能是为选定的CD34+细胞建立冷冻参数的第一步。
