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3D4/21
Tissue
肺
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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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

3D4/21 細胞系是一種廣泛應用於免疫反應與病毒感染研究的豬肺泡巨噬細胞模型。這些細胞對於研究豬隻的呼吸道感染(例如由豬流感病毒(SIV)和塞內卡病毒 A 型(SVA)所引起的感染)具有特別重要的價值。 由於源自肺泡巨噬細胞,3D4/21 細胞成為研究宿主與病原體相互作用的關鍵體外系統,重點在於探究豬隻對抗呼吸道病原體所啟動的免疫機制。

3D4/21 細胞常被用於探索感染過程中觸發的分子途徑。例如,在豬流感病毒(SIV)感染的背景下,研究顯示 3D4/21 細胞會透過表達各種細胞因子及病毒識別受體(如 RIG-I 和 TLR7)來啟動免疫反應。 此外,這些細胞在釐清不同 RNA 分子(包括 miRNA、lncRNA 和 circRNA)如何參與病毒感染期間的調控網絡方面發揮了關鍵作用,並為有助於抗病毒防禦的競爭性內源性 RNA(ceRNA)調控機制提供了見解。 這對於闡明由 H1N1 和 H3N2 等不同豬流感病毒株所觸發的免疫反應,尤為有用。

此外,3D4/21 細胞亦被用於研究自噬及其在感染控制中的作用。例如,在副豬嗜血桿菌(Haemophilus parasuis)感染期間,這些細胞會展現經由 AMPK 訊號傳導路徑調控的自噬反應。 這種自噬機制不僅有助於控制細菌入侵,也有助於理解如格拉瑟氏病(Glässer’s disease)等疾病的致病機制——該病症會影響處於壓力狀態下的豬隻。因此,3D4/21 細胞系在豬隻健康領域的免疫學與傳染病研究中,仍持續扮演著極具價值的工具角色。

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

02Classification

Organism
Pig
Sex
Unspecified
Growth properties
Adherent
Biosafety level
2

03Characteristics

Morphology
巨噬細胞
Cell type
肺泡巨噬細胞

04Identifiers

Citation
3D4/21(Cytion 型號 305357)
NCBI TaxID
9823
Cellosaurus
CVCL_0F14

05Expression and susceptibility

Viruses
轉化體:猿病毒 40 型(SV40)

06Handling and culture

Culture medium
RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% 的胎牛血清(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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

08Donor

Breed / subspecies
地方品種
Donor age
12 週