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BALB/3T3 clone A31
Tissue
胚胎
Synonyms
BALB/c 3T3 clone A31, Balb/c3T3, BALB/c 3T3, Balb/c 3T3, BALB/3T3, Balb/3T3-4-Cl31, 3T3 clone A31, BALB/3T3 cl. A31, BALB 3T3 clone A31, BALB/3T3 (clone A31), B/C3T3, 3T3-A31, 3T3(A31), A31, A31N

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BALB/3T3 clone A31

Cell line · Product number 305155

BALB/3T3 clone A31, a fibroblast cell line developed by S.A. Aaronson and G.T. Todaro in 1968, originates from disaggregated 14- to 17-day-old BALB/c mous...

US$395.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
  • CoA
1

01General information

Description

BALB/3T3 克隆 A31 是一種由 S.A. Aaronson 和 G.T. Todaro 於 1968 年開發的成纖維細胞系,源自經解聚處理的 14 至 17 天齡 BALB/c 小鼠胚胎。 該細胞系是細胞生物學研究中的基礎工具,尤其以其能支持病毒增殖以及易受致癌轉化影響而著稱。 這些細胞的特徵是呈紡錘形的成纖維細胞,可作為多潛能間質細胞。它們展現出根據微環境影響或培養條件分化為各種組織的潛力,凸顯了其在實驗模型中的多功能性。

BALB/3T3 克隆 A31 的細胞培養方法包括在達到匯合前進行反覆傳代,以盡量減少細胞間接觸,從而促進細胞分裂的接觸抑制、高稀釋度生長及低飽和密度等特性。 這些細胞的核型存在變異性,模態數為 78 條染色體,範圍介於 62 至 109 條之間,主要以端著絲點染色體或端著絲點染色體為主。 儘管偶有細胞遺傳學不穩定性的報告,BALB/3T3 A31 細胞仍維持非致瘤性狀態,但在半固體培養基中培養時則會展現致瘤性。 值得注意的是,它們極易受到 SV40 和小鼠肉瘤病毒等致癌性 DNA 病毒的轉化,且經檢測對外生性病毒(小鼠天花)呈陰性反應,這為病毒學和腫瘤學研究增添了另一層價值。

Tissue
胚胎
Synonyms
  • BALB/c 3T3 clone A31
  • Balb/c3T3
  • BALB/c 3T3
  • Balb/c 3T3
  • BALB/3T3
  • Balb/3T3-4-Cl31
  • 3T3 clone A31
  • BALB/3T3 cl. A31
  • BALB 3T3 clone A31
  • BALB/3T3 (clone A31)
  • B/C3T3
  • 3T3-A31
  • 3T3(A31)
  • A31
  • A31N

02Classification

Organism
Mouse
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
成纖維細胞
Tumourigenic
不,這些細胞在免疫抑制小鼠體內並未產生腫瘤,但在半固體培養基中確實形成了集落。

04Identifiers

Citation
BALB/3T3 克隆 A31(Cytion 型號 305155)
NCBI TaxID
10090
Cellosaurus
CVCL_0184

05Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
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
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
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
BALB/c
Donor age
胚胎,妊娠第14至17天