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Ana-1
Tissue
骨髓
Synonyms
ANA-1, ANA1

Documents

Ana-1

Cell line · Product number 305172

The Ana-1 cell line is a murine macrophage cell line derived from the ascites of a BALB/c mouse with a methylcholanthrene-induced tumor. This cell line is...

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1

01General information

Description

Ana-1 細胞系是一種源自罹患甲基膽蒽誘導性腫瘤之 BALB/c 小鼠腹水的鼠源巨噬細胞系。由於該細胞系具有產生多種細胞因子的能力,且在先天性免疫反應中扮演重要角色,因此在免疫學研究中被廣泛應用。 Ana-1 細胞具有吞噬活性,並能產生一氧化氮(NO),這對於針對病原體和腫瘤細胞的細胞毒性反應至關重要。

此外,Ana-1 細胞已被用作研究巨噬細胞活化及參與免疫反應之訊號傳導路徑的模型系統。該細胞會對脂多醣(LPS)刺激作出反應,透過上調促炎性細胞因子的表達來回應,因此非常適合用於探究發炎及宿主防禦系統的分子機制。 由於其與各種免疫學檢測方法的相容性,這些細胞對於藥物篩選以及理解巨噬細胞與免疫系統中其他細胞類型的相互作用,亦具有重要價值。

Tissue
骨髓
Synonyms
  • ANA-1
  • ANA1

02Classification

Organism
Mouse
Growth properties
Adherent and suspension
Biosafety level
1

03Characteristics

Morphology
巨噬細胞

04Identifiers

Citation
Ana-1(Cytion 型號 305172)
NCBI TaxID
10090
Cellosaurus
CVCL_0142

05Handling 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% 經熱滅活的胎牛血清
Dissociation reagent
Accutase
Subculturing
將懸浮細胞收集至 15 毫升試管中,並用不含鈣和鎂的 PBS 輕柔地洗滌貼壁細胞(T25 培養瓶使用 3-5 毫升,T75 培養瓶使用 5-10 毫升)。 加入 Accutase(T25 培養瓶使用 1–2 毫升,T75 培養瓶使用 2.5 毫升),確保完全覆蓋細胞層。讓細胞於室溫下孵育 10 分鐘。 孵育完成後,將懸浮細胞與貼壁細胞合併並進行離心。離心後,小心地將細胞沉澱重新懸浮,並將細胞懸液轉移至裝有新鮮培養基的新培養瓶中。
Seeding density
2 至 4 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
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.

06Quality 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。

07Donor

Breed / subspecies
C57BL/6