Bohbot A, Lioure B, Faradji A, Schmitt M, Cuillerot J M, Laplace A, Oberling F
Institut d'Hématologie et d'Immunologie, Hospices Civils, Faculté de Médecine, Université Louis Pasteur, Strasbourg, France.
Bone Marrow Transplant. 1996 Feb;17(2):259-64.
Autologous stem cell transplantation has become an important therapy in lymphoma, multiple myeloma and solid tumors. The rationale for the selection of CD34+ cells from peripheral blood or bone marrow progenitor cell collections is based on the observation that contaminating tumor cells can be depleted approximately 3 to 6 logs. This procedure may be limited because of lack of sufficient numbers of progenitor cells in the leukapheresis concentrates. The use of frozen/thawed peripheral blood mononuclear cell (PBMC) samples makes it possible to pool two or even more stem cell harvests collected at different time points to increase the total number of CD34+ progenitor cells. We report in this work the feasibility of frozen/thawed peripheral blood CD34+ positive cell selection, using the large-scale (Ceprate SC) and the lab-scale avidin-biotin immunoadsorption system (Ceprate LC). This procedure consists of a washing step and a positive selection step. Our results show that frozen/thawed CD34+ cells were obtained with a purity of 86.68 +/- 3.62%, a viability of 97.94 +/- 0.97% and a recovery of 91.85 +/- 10.84% (range 80 to 112%). The CFU-GM assays were performed in a methylcellulose based medium; 89.13 +/- 19.63 colonies were obtained for 10(3) cells plated. Two patients were grafted with peripheral blood CD34+ cells selected after freezing. Our clinical data show that these cells are capable of rapidly reconstituting hematopoiesis after high-dose chemotherapy.
自体干细胞移植已成为淋巴瘤、多发性骨髓瘤和实体瘤的重要治疗方法。从外周血或骨髓祖细胞采集中选择CD34+细胞的基本原理是基于这样的观察结果：污染的肿瘤细胞可被清除约3至6个对数级。由于白细胞分离浓缩物中祖细胞数量不足，该方法可能受到限制。使用冻融外周血单个核细胞（PBMC）样本可以汇集在不同时间点采集的两个甚至更多干细胞收获物，以增加CD34+祖细胞的总数。我们在这项工作中报告了使用大规模（Ceprate SC）和实验室规模的抗生物素蛋白-生物素免疫吸附系统（Ceprate LC）进行冻融外周血CD34+阳性细胞分选的可行性。该过程包括洗涤步骤和阳性分选步骤。我们的结果表明，获得的冻融CD34+细胞纯度为86.68±3.62%，活力为97.94±0.97%，回收率为91.85±10.84%（范围80%至112%）。CFU-GM测定在基于甲基纤维素的培养基中进行；接种10(3)个细胞可获得89.13±19.63个集落。两名患者接受了冷冻后分选的外周血CD34+细胞移植。我们的临床数据表明，这些细胞能够在大剂量化疗后迅速重建造血功能。
