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Optimized simple and affordable procedure for differentiation of monocyte-derived dendritic cells from LRF: An accessible and valid alternative biological source
Experimental Cell Research ( IF 3.3 ) Pub Date : 2021-07-30 , DOI: 10.1016/j.yexcr.2021.112754
Maryam Valizadeh 1 , Ali Akbar Purfathollah 1 , Reza Raoofian 2 , Afrooz Homayoonfar 1 , Mohammad Moazzeni 1
Affiliation  

Dendritic cells are one of the most popular immune cells, which plays a remarkable role in both immunotherapy and tolerance induction. Due to unwanted side effects of leukocyte presence in donated blood, the policy of blood service is the pre-storage reduction of leukocytes, which today, filtration is the most common method for this purpose. The filtration method has led to diminished access to Buffy coat as a generally used conventional source of biological cells. We developed a simple, affordable, and reproducible method for dendritic cell differentiation from filter-derived monocytes and, the results of the filter study were compared with differentiated DCs from the conventional buffy coat-derived monocytes. The Monocytes were recovered from leukoreduction filter using an optimized protocol with supplemented PBS buffer. Following the adhesion method, CD14+ Monocyte-enriched population with the purity of 94 % was obtained. After cytokine stimulation over a 6-day period and maturation induction by LPS, differentiated DCs were evaluated for morphology, surface markers (CD86, CD40, CD83 and, HLA-DR), antigen uptake potency and IL-12 secretion. Analysis and comparison of the results represented no significant difference between the two groups. Accordingly, we conclude that leukoreduction filters could be introduced as a reliable and research-grade source of monocyte for DC generation in biological research.



中文翻译:

从 LRF 中分化出单核细胞衍生的树突细胞的优化简单且经济实惠的程序:一种可获得且有效的替代生物来源

树突状细胞是最受欢迎的免疫细胞之一,在免疫治疗和耐受诱导中都发挥着显着的作用。由于捐献血液中存在白细胞会产生不必要的副作用,血液服务的政策是预先减少白细胞的储存,今天,过滤是用于此目的的最常用方法。过滤方法导致减少了对作为普遍使用的常规生物细胞来源的白膜层的访问。我们开发了一种简单、经济且可重复的方法,用于从过滤器衍生的单核细胞分化树突细胞,并将过滤器研究的结果与来自传统血沉棕黄层衍生单核细胞的分化 DC 进行比较。使用带有补充 PBS 缓冲液的优化方案从白细胞减少过滤器中回收单核细胞。+获得了纯度为 94% 的富含单核细胞的群体。经过 6 天的细胞因子刺激和 LPS 诱导成熟后,对分化的 DC 进行形态学、表面标志物(CD86、CD40、CD83 和 HLA-DR)、抗原吸收能力和 IL-12 分泌进行评估。结果分析比较,两组间无显着差异。因此,我们得出结论,白细胞减少过滤器可以作为一种可靠的研究级单核细胞来源,用于生物研究中的 DC 生成。

更新日期:2021-08-15
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