Automated generation of immature dendritic cells in a single-use system.
Automated generation of immature dendritic cells in a single-use system.
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DOI：
10.1016/j.jim.2018.03.010
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发表时间：
2018-06
影响因子：
2.2
通讯作者：
Murthy SK
中科院分区：
文献类型：
--
作者：
Kozbial A;Bhandary L;Collier BB;Eickhoff CS;Hoft DF;Murthy SK
树突状细胞 (DC) 是研究人类反应不可或缺的一部分，这对于针对癌症和传染病的保护性免疫以及预防自身免疫和移植排斥非常重要。这些细胞也是癌症和传染病个性化疫苗的关键要素。尽管树突状细胞在临床和基础研究中都发挥着重要作用，但从个体中获取这些细胞的方法仍然相对不发达且效率低下。 DC 在血液中的浓度非常低 (<1%)，因此它们必须由单核细胞产生，而目前 DC 产生的方法涉及静态培养和用培养基中所含细胞因子刺激的费力过程。在此，我们描述了一种自动化流体系统 MicroDEN，它允许利用分化培养基的连续灌注将单核细胞分化为未成熟的 DC（iDC）。与当前离体单核细胞分化相关的手动步骤大大减少，并且通过使用封闭的盒和管道网络确保了无菌环境。对生成的 iDC 进行基准表型分析以及同种异体 T 细胞增殖和同基因抗原特异性功能测定。 MicroDEN 生成的 iDC 在表型和功能上与孔板生成的 iDC 相似，从而证明了在需要 DC 的广泛应用中利用 MicroDEN 的可行性。
Dendritic cells (DCs) are an indispensable part of studying human responses that are important for protective immunity against cancer and infectious diseases as well as prevention of autoimmunity and transplant rejection. These cells are also key elements of personalized vaccines for cancer and infectious diseases. Despite the vital role of DCs in both clinical and basic research contexts, methods for obtaining these cells from individuals remains a comparatively under-developed and inefficient process. DCs are present in very low concentrations (<1%) in blood, thus they must be generated from monocytes and the current methodology in DC generation involves a laborious process of static culture and stimulation with cytokines contained in culture medium. Herein, we describe an automated fluidic system, MicroDEN, that allows for differentiation of monocytes into immature-DCs (iDCs) utilizing continuous perfusion of differentiation media. Manual steps associated with current ex vivo monocyte differentiation are vastly reduced and an aseptic environment is ensured by the use of an enclosed cartridge and tubing network. Benchmark phenotyping was performed on the generated iDCs along with allogeneic T-cell proliferation and syngeneic antigen-specific functional assays. MicroDEN generated iDCs were phenotypically and functionally similar to well plate generated iDCs, thereby demonstrating the feasibility of utilizing MicroDEN in the broad range of applications requiring DCs.
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影响因子：
5.8
作者：
Phuphanich, Surasak;Wheeler, Christopher J.;Rudnick, Jeremy D.;Mazer, Mia;Wang, HongQian;Nuno, Miriam A.;Richardson, Jaime E.;Fan, Xuemo;Ji, Jianfei;Chu, Ray M.;Bender, James G.;Hawkins, Elma S.;Patil, Chirag G.;Black, Keith L.;Yu, John S.
通讯作者：
Yu, John S.
影响因子：
20.3
作者：
Kochenderfer, James N.;Wilson, Wyndham H.;Rosenberg, Steven A.
通讯作者：
Rosenberg, Steven A.
影响因子：
15.9
作者：
Liu, Quan;Rojas-Canales, Darling M.;Morelli, Adrian E.
通讯作者：
Morelli, Adrian E.
影响因子：
4.4
作者：
Dumortier, H;van Mierlo, GJD;Melief, CJM
通讯作者：
Melief, CJM
影响因子：
4.6
作者：
Fishman, Mayer
通讯作者：
Fishman, Mayer