Centre de Thérapie Cellulaire, Institut Paoli-Calmettes, Centre Régional de Lutte Contre le Cancer Provence-Alpes-Côte d'Azur, 232 Boulevard de Ste Marguerite, BP 156. 13273 Marseille Cedex 9, France.
Cytotherapy. 1999;1(6):469-77.
Enrichment in mononuclear cells (MNC) is a prerequisite for CD34(+) cell selection from BM allografts. However, centrifugation over a density gradient (Ficoll), does not comply with good manufacturing practice (GMP), because Ficoll is not a clinical grade reagent. We evaluated the COBE Spectra cell separator as an alternative and evaluated the recovery of MNC and CD34(+) cells. CD34(+) cell selection was then performed using either the Nexell Isolex 300i or the CEPRATE SC (CellPro) device.
20 allogeneic BM grafts were processed, with an initial enrichment of MNC using the COBE Spectra processor, followed by immunoselection of CD34(+) cells (13 Isolex 300i and seven CEPRATE SC). We evaluated total nucleated cells (TNC), MNC and CD34(+) cells recoveries when using COBE Spectra and compared CD34(+) cells recovery and T-cell depletion when using Isolex 300i or CEPRATE SC.
Median recoveries of the product using the COBE Spectra were 31.2% (18.1-68.9%) for TNC, 76.9% (29.5-148.1%) for MNC and 79.6% (60.9-191%) for CD34(+) cells. Median recoveries of CD34(+) cells were 39.4% (15.1-75.8%) and 36.7% (26.2-64.3%) with the Isolex 300i and CEPRATE SC devices, respectively. CD34(+) purity was 85% (64.4-94.5%) and 84.4% (66.8-86.8%). Colony-forming unit granulocyte/macrophage (CFU-GM) recoveries were 10.9% (0.2-152%) and 36% (0.0-67.3%). Median log depletion for T-cell numbers, B-cell numbers and natural killer (NK) cell numbers were 2.8 log (2.1-3.8), 2.0log (0.95-3.1) and 3.1log (2.40-4.8), respectively.
Processing with COBE Spectra produces a cell population that is suitable for CD34(+) cell selection. The Isolex 300i and CEPRATE SC devices demonstrate similar performances. The two-step procedure allows allo-BMT in patients with high risk of GvHD.
从骨髓异基因移植物中选择 CD34(+)细胞需要富集单个核细胞 (MNC)。然而，使用密度梯度（Ficoll）离心不能符合良好生产规范（GMP），因为 Ficoll 不是临床级别的试剂。我们评估了 COBE Spectra 细胞分离机作为替代方法，并评估了 MNC 和 CD34(+)细胞的回收情况。然后使用 Nexell Isolex 300i 或 CEPRATE SC（CellPro）设备进行 CD34(+)细胞选择。
处理了 20 例异基因骨髓移植物，使用 COBE Spectra 处理器对 MNC 进行初始富集，然后对 CD34(+)细胞进行免疫选择（13 例 Isolex 300i 和 7 例 CEPRATE SC）。我们评估了使用 COBE Spectra 时的总核细胞 (TNC)、MNC 和 CD34(+)细胞回收率，并比较了使用 Isolex 300i 或 CEPRATE SC 时的 CD34(+)细胞回收率和 T 细胞耗竭情况。
使用 COBE Spectra 时，产品的中位数回收率为 TNC 31.2%（18.1-68.9%），MNC 76.9%（29.5-148.1%）和 CD34(+)细胞 79.6%（60.9-191%）。使用 Isolex 300i 和 CEPRATE SC 设备时，CD34(+)细胞的中位数回收率分别为 39.4%（15.1-75.8%）和 36.7%（26.2-64.3%）。CD34(+)纯度为 85%（64.4-94.5%）和 84.4%（66.8-86.8%）。粒-巨噬细胞集落形成单位 (CFU-GM)回收率为 10.9%（0.2-152%）和 36%（0.0-67.3%）。T 细胞数量、B 细胞数量和自然杀伤 (NK) 细胞数量的中位数对数耗竭分别为 2.8 对数（2.1-3.8）、2.0 对数（0.95-3.1）和 3.1 对数（2.40-4.8）。
使用 COBE Spectra 进行处理可产生适合 CD34(+)细胞选择的细胞群。Isolex 300i 和 CEPRATE SC 设备表现相似。两步法可使高危移植物抗宿主病患者接受同种异体 BMT。
