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Generation of potent porcine monocyte-derived dendritic cells (MoDCs) by modified culture protocol

机译:通过修改的培养方案生成有效的猪单核细胞衍生树突状细胞(MoDC)

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In vitro derivation of dendritic cells (DCs) is an alternative approach to overcome the low frequency of primary DCs and the difficulty of isolation techniques for studying DC immunobiology. To date, the conventional culture protocol of porcine monocyte-derived DCs (MoDCs) has been widely used. However, this protocol is not practical due to the requirement of a substantial number of blood monocytes, and the process often interferes with DC maturation. To improve in vitro porcine MoDC generation, we modified the previous conventional DC generation protocol, based on the human and mouse primary DC culture system, and compared phenotypic and functional features of MoDCs derived from the modified protocol to the conventional protocol. The modified protocol consumed fewer monocytes but generated higher CD1(+) cells with DC-like morphology and the ability of maturation. In addition, MoDCs from the modified protocol exhibited increased antigen uptake and IFN-gamma production in response to LPS stimulation. Our findings indicate that the modified protocol is expedient and reliable for generating potent MoDCs that substitute for primary DCs. This will be a valuable platform for future research in antigen delivery, vaccines and immunotherapy in pigs, as well as relevant veterinary species. (C) 2016 Elsevier B.V. All rights reserved.
机译:树突状细胞(DC)的体外衍生是克服原代DC频率低和研究DC免疫生物学的分离技术困难的另一种方法。迄今为止,猪单核细胞来源的DC(MoDC)的常规培养方案已被广泛使用。但是,由于需要大量的血液单核细胞,因此该方案不切实际,并且该过程通常会干扰DC成熟。为了改善体外猪MoDC的生成,我们基于人和小鼠的主要DC培养系统修改了以前的常规DC生成协议,并将从修改后的协议衍生的MoDC的表型和功能特征与常规协议进行了比较。修改后的协议消耗较少的单核细胞,但生成具有DC样形态和成熟能力的较高CD1(+)细胞。另外,来自改良方案的MoDCs响应LPS刺激而表现出增加的抗原摄取和IFN-γ产生。我们的发现表明,修改后的协议对于生成可替代主DC的有效MoDC既方便又可靠。这将是未来在猪以及相关兽医物种中进行抗原递送,疫苗和免疫疗法研究的宝贵平台。 (C)2016 Elsevier B.V.保留所有权利。

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