Genomic Sciences- R&D, GSK, Collegeville, PA, USA.
Department of Pathology, New York University School of Medicine, New York, NY, USA.
Methods Mol Biol. 2023;2618:109-119. doi: 10.1007/978-1-0716-2938-3_8.
Dendritic cells (DCs) comprise a heterogeneous population of antigen (Ag)-presenting cells that play a critical role in both innate and adaptive immunity. DCs orchestrate protective responses against pathogens and tumors while mediating tolerance to host tissues. Evolutionary conservation between species has allowed the successful use of murine models to identify and characterize DC types and functions relevant to human health. Among DCs, type 1 classical DCs (cDC1) are uniquely capable of inducing antitumor responses and therefore present a promising therapeutic target. However, the rarity of DCs, particularly cDC1, limits the number of cells that can be isolated for study. Despite significant effort, progress in the field has been hampered by inadequate methods to produce large quantities of functionally mature DCs in vitro. To overcome this challenge, we developed a culture system in which mouse primary bone marrow cells are cocultured with OP9 stromal cells expressing Notch ligand Delta-like 1 (OP9-DL1) to produce CD8α DEC205 XCR1 cDC1 (Notch cDC1). This novel method provides a valuable tool to facilitate the generation of unlimited cDC1 for functional studies and translational applications such as antitumor vaccination and immunotherapy.
树突状细胞 (DCs) 是一类具有异质性的抗原 (Ag) 呈递细胞，在固有和适应性免疫中发挥着关键作用。DCs 协调针对病原体和肿瘤的保护反应，同时介导对宿主组织的耐受。物种间的进化保守性使得成功地使用小鼠模型来鉴定和表征与人类健康相关的 DC 类型和功能成为可能。在 DCs 中，1 型经典 DC (cDC1) 具有独特的诱导抗肿瘤反应的能力，因此是一个很有前途的治疗靶点。然而，DCs 的稀有性，特别是 cDC1 的稀有性，限制了可用于研究的细胞数量。尽管付出了巨大的努力，但由于缺乏在体外大量产生功能成熟 DCs 的方法，该领域的进展受到了阻碍。为了克服这一挑战，我们开发了一种培养系统，其中将小鼠原代骨髓细胞与表达 Notch 配体 Delta-like 1 (OP9-DL1) 的 OP9 基质细胞共培养，以产生 CD8α DEC205 XCR1 cDC1 (Notch cDC1)。这种新方法为功能研究和转化应用（如抗肿瘤疫苗接种和免疫治疗）提供了一种有价值的工具，可用于产生无限量的 cDC1。
