Kim Sinyoung, Kim Hyun Ok, Baek Eun-Jung, Choi Youjeong, Kim Han-Soo, Lee Min-Geul
Department of Laboratory Medicine, Yonsei Cell Therapy Center, Department of Dermatology, Yonsei University College of Medicine, Seoul, Korea.
Transfusion. 2007 Dec;47(12):2290-6. doi: 10.1111/j.1537-2995.2007.01470.x. Epub 2007 Aug 30.
Dendritic cells (DCs), used in clinical trials for cancer immunotherapy, require processing on an expanded scale to conform to current good manufacturing practice guidelines. This study evaluated a large-scale monocyte enrichment procedure with a commercially available cell separator (Elutra, Gambro BCT) and analyzed the capacity of enriched monocytes to differentiate into DCs.
Mononuclear cells were collected in two patients with malignant melanoma and seven healthy donors by leukapheresis. Continuous-counterflow elutriation with the Elutra was performed to enrich and purify monocytes from leukapheresis products. Purity and recovery of enriched monocytes were analyzed by flow cytometry. DCs were generated from the elutriated monocytes and characterized by phenotypic surface marker and stimulatory capacity in an allogeneic mixed lymphocyte reaction.
In the leukapheresis products, the total MNC count was 7.3 x 10(9) +/- 0.7 x 10(9) and the mean percentage of CD14+ monocytes was 16.5 +/- 3.8 percent, which increased to 68.9 +/- 7.4 percent after elutriation with the Elutra. The mean monocyte recovery was 94.3 percent. Elutriated monocytes were successfully cultured into phenotypically and functionally mature DCs.
These results indicate that the Elutra cell separator allows for fast and easy enrichment of monocytes within a closed system. Furthermore, these monocytes can be differentiated into functionally mature DCs. Compared to plastic adherence and immunomagnetic selection methods, the elutriation procedure is inexpensive, efficient, and very effective.
用于癌症免疫治疗临床试验的树突状细胞（DCs）需要大规模处理以符合现行良好生产规范指南。本研究评估了使用市售细胞分离器（Elutra，甘布罗BCT公司）进行大规模单核细胞富集程序，并分析了富集单核细胞分化为DCs的能力。
通过白细胞分离术收集了2例恶性黑色素瘤患者和7名健康供者的单核细胞。使用Elutra进行连续逆流淘析，以从白细胞分离产物中富集和纯化单核细胞。通过流式细胞术分析富集单核细胞的纯度和回收率。从淘析后的单核细胞生成DCs，并通过表型表面标志物和同种异体混合淋巴细胞反应中的刺激能力进行表征。
在白细胞分离产物中，总单核细胞计数为7.3×10⁹±0.7×10⁹，CD14⁺单核细胞的平均百分比为16.5±3.8%，使用Elutra淘析后增加至68.9±7.4%。单核细胞的平均回收率为94.3%。淘析后的单核细胞成功培养为表型和功能成熟的DCs。
这些结果表明，Elutra细胞分离器可在封闭系统中快速简便地富集单核细胞。此外，这些单核细胞可分化为功能成熟的DCs。与塑料贴壁和免疫磁选方法相比，淘析程序价格低廉、高效且非常有效。
