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3D4/21
Tissue
肺部
Synonyms
IPAM 3D4/21, IPAM-WT

Documents

3D4/21

Cell line · Product number 305357

The 3D4/21 cell line is a porcine alveolar macrophage model extensively used in studies involving immune responses and viral infections. These cells are p...

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1

01General information

Description

3D4/21细胞系是一种猪肺泡巨噬细胞模型,广泛应用于免疫应答和病毒感染相关研究。这些细胞对于研究猪的呼吸道感染(例如由猪流感病毒(SIV)和塞内卡病毒A型(SVA)引起的感染)具有特别重要的价值。 由于源自肺泡巨噬细胞,3D4/21细胞作为研究宿主-病原体相互作用的关键体外系统,重点探讨猪针对呼吸道病原体所启动的免疫机制。

3D4/21细胞常被用于探索感染过程中触发的分子通路。例如,在SIV感染背景下,研究表明3D4/21细胞通过表达多种细胞因子以及RIG-I和TLR7等病毒识别受体来启动免疫应答。 此外,这些细胞在阐明包括miRNA、lncRNA和circRNA在内的不同RNA分子如何参与病毒感染期间的调控网络方面发挥了关键作用,从而深入揭示了有助于抗病毒防御的竞争性内源性RNA(ceRNA)调控机制。 这在阐明不同猪流感病毒株(如H1N1和H3N2)所触发的免疫应答方面尤为有用。

此外,3D4/21细胞还被用于研究自噬及其在感染控制中的作用。例如,在副猪嗜血杆菌感染期间,这些细胞会表现出通过AMPK信号通路调节的自噬反应。 这种自噬机制不仅有助于控制细菌入侵,还有助于理解格拉瑟病等疾病的发病机制——该病是一种影响处于应激状态下的猪的疾病。因此,3D4/21细胞系在猪健康领域的免疫学和传染病研究中,仍然是一个有价值的工具。

Tissue
肺部
Synonyms
  • IPAM 3D4/21
  • IPAM-WT

02Classification

Organism
Pig
Sex
Unspecified
Growth properties
Adherent
Biosafety level
2

03Characteristics

Morphology
巨噬细胞
Cell type
肺泡巨噬细胞

04Identifiers

Citation
3D4/21(Cytion 产品编号 305357)
NCBI TaxID
9823
Cellosaurus
CVCL_0F14

05Expression and susceptibility

Viruses
转化体: Simian virus 40 (SV40)

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
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Seeding density
1-3×10⁴/cm²
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
兰德瑞斯
Donor age
12周