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DC2.4
Tissue
骨髓
Synonyms
DC 2.4

Documents

DC2.4

Cell line · Product number 305515

The DC2.4 cell line is an immortalized mouse dendritic cell line that originates from bone marrow. It is commonly used to study dendritic cell (DC) biolog...

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01General information

Description

DC2.4 细胞系是一种源于骨髓的永生化小鼠树突状细胞系。它通常用于研究树突状细胞(DC)生物学、免疫反应和免疫疗法的开发。DC2.4 细胞的特点是作为抗原递呈细胞(APC),并表达树突状细胞的典型表面标记,如 CD11c 和 MHC I 类分子。然而,在标准培养条件下,它们表现出不成熟的表型,MHC II 类和成本刺激分子(如 CD40 和 CD80)的表达量较低。这使它们成为研究直流细胞成熟及其后续免疫功能所需的机制和刺激的有用工具。

研究表明,特定刺激可诱导 DC2.4 细胞成熟。值得注意的是,暴露于γ干扰素(IFN-γ)会导致 MHC II 类、CD40、CD80 和 CCR7 的显著上调,以及细胞因子分泌的增加,包括 IL-6、IL-12 和 TNF-α。已证明IFN-γ成熟的DC2.4细胞能在体外和体内有效激活CD8+细胞毒性T细胞,增强抗肿瘤免疫力。例如,经 IFN-γ 处理、抗原刺激的 DC2.4 细胞已被证明能诱导强大的 CD8+ T 细胞反应,并在小鼠模型中提供保护性抗肿瘤效应。这凸显了该细胞系在癌症免疫疗法研究和疫苗开发中的实用性。

此外,DC2.4 细胞还被用来研究宿主与病原体之间的相互作用,因为它们对各种免疫挑战的反应可以模拟先天性免疫系统激活的各个方面。对 DC2.4 细胞外泌体 miRNA 图谱的分析,尤其是在感染弓形虫等病原体时的分析,有助于深入了解树突状细胞信号传导和免疫通讯的分子机制。外泌体 miRNA 在感染反应中的不同表达表明了它们在调节宿主免疫中的潜在作用,并突出了 DC2.4 在外泌体和基于 RNA 的免疫研究中的实用性。

Tissue
骨髓
Synonyms
  • DC 2.4

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine dendritic cell line (DC2.4) contains retroviral constructs encoding murine GM-CSF, v-myc and v-raf introduced by transduction, supporting transformation and growth. The inserts are stably present in the dendritic cell-derived line. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Cell type
树突状细胞

04Identifiers

Citation
DC2.4(Cytion 目录编号 305515)
NCBI TaxID
10090
Cellosaurus
CVCL_J409

05Expression and susceptibility

Viruses
转基因重组逆转录病毒 J2

06Handling and culture

Culture medium
RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
向培养基中加入10% FBS、1% NEAA 和 10mM HEPES
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。

Freeze medium
我们使用完全生长培养基(包括 FBS)+10% DMSO 或 CM-1(Cytion 商品目录编号 800100)作为冷冻保存培养基,以获得足够的解冻后存活率,CM-1(Cytion 商品目录编号 800100)含有优化的渗透保护剂和代谢稳定剂,可提高恢复能力并减少冷冻引起的应激。
Storage conditions

如需长期保存,可将样品瓶放入气相液氮中,温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运,并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后,应立即检查容器,并立即将小瓶转移到适当的储藏室。

08Donor

Breed / subspecies
C57BL/6
Donor age
未说明