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树突状细胞生成与成熟过程中的培养基及蛋白质补充剂

Culture medium and protein supplementation in the generation and maturation of dendritic cells.

作者信息

Royer P-J, Tanguy-Royer S, Ebstein F, Sapede C, Simon T, Barbieux I, Oger R, Gregoire M

机构信息

INSERM U601, Institut de Biologie, Nantes, France.

出版信息

Scand J Immunol. 2006 Jun;63(6):401-9. doi: 10.1111/j.1365-3083.2006.001757.x.

Abstract

Dendritic cells (DC) are powerful antigen-presenting cells that have drawn many attentions due to the recent development of anti-cancer vaccines. Clinical grade production of monocyte-derived DC (Mo-DC) is extensively studied, and many efforts are made to develop and improve clinical standard operating procedures. Most of the parameters involved, such as the cytokines and maturation agents, have been widely assessed. However, very few are investigated about how culture medium and additional protein components affect DC yield, viability and maturation. Thus, our study aimed to compare the impact of standard culture medium on Mo-DC differentiation and maturation. Commercially available media for hematopoietic cell culture as well as different protein supplementations, that is foetal calf serum (FCS), autologous plasma (AP), human serum (HS) and human serum albumin (HSA) were tested. Culture yields, cell viability and DC maturation were investigated. Differentiation yields were similar between the conditions used. However, we evidenced significant differences in terms of cytotoxicity and DC maturation (phenotypic and functional). This underscores the importance of defining culture medium composition in clinical standard operating procedures to insure quality control, and also when preparing DC for experimental uses.

摘要

树突状细胞(DC)是强大的抗原呈递细胞,由于抗癌疫苗的最新进展而备受关注。单核细胞衍生的DC(Mo-DC)的临床级生产已得到广泛研究,并且人们为制定和完善临床标准操作程序付出了诸多努力。所涉及的大多数参数,如细胞因子和成熟剂,都已得到广泛评估。然而,关于培养基和其他蛋白质成分如何影响DC产量、活力和成熟的研究却很少。因此,我们的研究旨在比较标准培养基对Mo-DC分化和成熟的影响。我们测试了市售的造血细胞培养基以及不同的蛋白质补充剂,即胎牛血清(FCS)、自体血浆(AP)、人血清(HS)和人血清白蛋白(HSA)。研究了培养产量、细胞活力和DC成熟情况。所用条件之间的分化产量相似。然而,我们证明在细胞毒性和DC成熟(表型和功能)方面存在显著差异。这凸显了在临床标准操作程序中确定培养基成分以确保质量控制的重要性,以及在为实验用途制备DC时的重要性。

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