Rapid debulking and CD34 enrichment of filgrastim-mobilized peripheral blood stem cells by semiautomated density gradient centrifugation in a closed system.

Rapid debulking and CD34 enrichment of filgrastim-mobilized peripheral blood stem cells by semiautomated density gradient centrifugation in a closed system.
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在封闭系统中通过半自动密度梯度离心对非格司亭动员的外周血干细胞进行快速减灭和 CD34 富集。

DOI:
10.1089/scd.1.1996.5.497
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发表时间:
1996
期刊:
Journal of hematotherapy
影响因子:
--
通讯作者:
Martin Körbling
Martin Körbling
中科院分区:
--
文献类型:
--
作者:
Donna Przepiorka;Peter VAN VLASSELAER;Lap Huynh;April Durett;Phylisha Agbor;J. Lauppe;Frank Valone;R. Champlin;Martin Körbling

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非格司亭动员的外周血祖细胞(PBPC)用于恶性肿瘤清髓治疗后的造血重建,但单次血浆分离术中收集的大量细胞经常会带来储存或进一步处理的问题。我们评估了 CD34 浮力密度溶液-PBPC(一种超轻密度胶体二氧化硅悬浮液)在半自动系统中用于 PBPC 制剂中 CD34+ 细胞减灭和富集的用途。使用 COBE Spectra 从四名经非格司亭治疗的正常供体中收集细胞。使用 COBE 2991 使用 Plasma-Lyte A 和 DNase 5 U/ml 进行分离程序。处理和洗涤后,有核细胞回收率为 26%,CD34+ 细胞富集 2.6 倍,CD34+ 细胞回收率 68%,CFU-GM 回收率 88%,BFU-E 回收率 73%,CD3+ 细胞消耗 1 个对数, CD56+ 细胞减少 0.5 log,CD19+ 细胞减少 1 log。这些结果与通过在管中离心在 CD34 浮力密度溶液-PBPC 上分离 PBPC 时获得的结果没有显着差异。使用 CD34 浮力密度溶液-PBPC,可以使用根据良好生产规范 (GMP) 生产的试剂,在半自动封闭系统中快速富集 PBPC 的单核制剂中的 CD34+ 细胞并耗尽淋巴细胞。
Filgrastim-mobilized peripheral blood progenitor cells (PBPC) are used for hematopoietic reconstitution after myeloablative therapy for malignancies, but the large number of cells collected in a single apheresis frequently presents a problem for storage or further processing. We have evaluated the use of CD34 Buoyant Density Solution-PBPC, an ultralight-density colloidal silica suspension, for debulking and enrichment of CD34+ cells in PBPC preparations in a semiautomated system. Cells were collected from four filgrastim-treated normal donors using the COBE Spectra. The separation procedure was carried out with Plasma-Lyte A and DNase 5 U/ml using the COBE 2991. Following processing and washing, there was a 26% recovery of nucleated cells, 2.6-fold enrichment of CD34+ cells, 68% recovery of CD34+ cells, 88% recovery of CFU-GM, 73% recovery of BFU-E, 1 log depletion of CD3+ cells, 0.5 log depletion of CD56+ cells, and 1 log depletion of CD19+ cells. These results were not significantly different from those obtained when PBPC were separated over CD34 Buoyant Density Solution-PBPC by centrifugation in tubes. Using CD34 Buoyant Density Solution-PBPC, mononuclear preparations of PBPC can be enriched rapidly for CD34+ cells and depleted of lymphocytes in a semiautomated closed system using reagents produced according to good manufacturing practice (GMP).
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