Tianjin Institute of Urology, The Second Hospital of Tianjin Medical University.
Department of Pathology, The Second Hospital of Tianjin Medical University.
J Vis Exp. 2022 Jul 1(185). doi: 10.3791/63125.
The demand for dendritic cells (DCs) is gradually increasing as immunology research advances. However, DCs are rare in all tissues. The traditional method for isolating DCs primarily involves inducing bone marrow (BM) differentiation into DCs by injecting large doses (>10 ng/mL) of granulocyte-macrophage colony-stimulating factor/interleukin-4 (GM-CSF/IL-4), making the procedure complex and expensive. In this protocol, using all BM cells cultured in 10 ng/mL GM-CSF/IL-4 medium, after 3-4 half-culture exchanges, up to 2.7 x 10 CD11c cells (DCs) per mouse (two femurs) were harvested with a purity of 80%-95%. After 10 days in culture, the expression of CD11c, CD80, and MHC II increased, whereas the number of cells decreased. The number of cells peaked after 7 days of culture. Moreover, this method only took 10 min to harvest all bone marrow cells, and a high number of DCs were obtained after 1 week of culture.
随着免疫学研究的进展，对树突状细胞（DCs）的需求逐渐增加。然而，所有组织中的 DCs 都很少。传统的分离 DCs 的方法主要通过注射大剂量（>10ng/mL）的粒细胞-巨噬细胞集落刺激因子/白细胞介素-4（GM-CSF/IL-4）来诱导骨髓（BM）分化为 DCs，操作复杂且昂贵。在本方案中，使用在 10ng/mL GM-CSF/IL-4 培养基中培养的所有 BM 细胞，经过 3-4 次半培养交换，每只小鼠（两根股骨）可收获多达 2.7×10 CD11c 细胞（DCs），纯度为 80%-95%。培养 10 天后，CD11c、CD80 和 MHC II 的表达增加，而细胞数量减少。培养 7 天后细胞数量达到峰值。此外，该方法仅需 10 分钟即可收获所有骨髓细胞，培养 1 周后即可获得大量 DCs。
