Efficient Generation of Bone Marrow-Derived Murine Dendritic Cells: A Protocol ‎Optimization Study.

IF 1.1 4区 医学 Q4 IMMUNOLOGY
Marziyeh Mousazadeh, Maryam Nikkhah, Negar Seyed, Sima Rafati, Saman Hosseinkhani
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引用次数: 0

Abstract

Background: Dendritic cells (DCs) are the most potent antigen-presenting cells, playing a central role in T cell activation and the stimulation of B cell antibody production. Owing to their immunological significance, DCs have been extensively explored for applications in cancer immunotherapy, dendritic cell-based vaccines, and strategies to overcome the immunosuppressive tumor microenvironment.

Objective: Given the broad applications of DCs and the need for efficient generation protocols, the present study aimed to optimize key parameters influencing the production of bone marrow-derived dendritic cells (BMDCs).

Methods: Variables such as mouse strain, cytokine concentrations, culture plate type, differentiation duration, incubation temperature, cell seeding density, and media replacement intervals were evaluated. CD11c expression was used as the primary marker to quantify BMDCs, while CD80 and CD86 were used as indicators of maturation status.

Results: The optimized protocol yielded 60-70% CD11c+ BMDCs and 70-80% CD80/CD86 expression within 5 days.

Conclusion: This robust and highly reproducible protocol offers valuable applications for in vitro dendritic cell generation studies.

高效生成骨髓来源的小鼠树突状细胞:一项方案优化研究。
背景:树突状细胞(dc)是最有效的抗原呈递细胞,在T细胞活化和刺激B细胞抗体产生中起着核心作用。由于其免疫学意义,树突状细胞已被广泛用于癌症免疫治疗、基于树突状细胞的疫苗和克服免疫抑制肿瘤微环境的策略。目的:考虑到骨髓源性树突状细胞的广泛应用和高效生成方案的需求,本研究旨在优化影响骨髓源性树突状细胞(bmdc)生成的关键参数。方法:对小鼠品系、细胞因子浓度、培养板类型、分化时间、孵育温度、细胞播种密度、培养基更换间隔等变量进行评价。CD11c表达被用作量化BMDCs的主要标记,而CD80和CD86被用作成熟状态的指标。结果:优化后的方案在5天内CD11c+ BMDCs表达率为60-70%,CD80/CD86表达率为70-80%。结论:该方法具有高度可重复性,为体外树突状细胞生成研究提供了有价值的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Iranian Journal of Immunology
Iranian Journal of Immunology Medicine-Immunology and Allergy
CiteScore
1.60
自引率
0.00%
发文量
50
审稿时长
12 weeks
期刊介绍: The Iranian Journal of Immunology (I.J.I) is an internationally disseminated peer-reviewed publication and publishes a broad range of experimental and theoretical studies concerned with all aspects of immunology.
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