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LC-540
Tissue
睪丸
Disease
腺瘤
Synonyms
LC540, LC 540

Documents

LC-540

Cell line · Product number 500262

The LC-540 cell line is an adherent cell model derived from an adult male Fischer rat. Known for its robust growth properties, this cell line has a modal ...

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01General information

Description

LC-540 細胞系是一種源自成年雄性費雪鼠的貼壁細胞模型。該細胞系以其強健的生長特性著稱,其模態染色體數為 42,核型範圍為 40 至 43。 約 21% 的細胞呈現非整倍體現象,儘管尚未有其他結構異常的報告,這顯示其基因組特徵相對穩定。

LC-540 細胞具有致瘤性,當植入大鼠體內時能形成腫瘤。此特性使其在受控的體外環境中,對於研究致癌機制與腫瘤生物學具有特別重要的價值。 此外,這些細胞易受多種病毒感染,包括單純疱疹病毒、牛痘病毒、水疱性口炎病毒及人類小兒麻痺病毒 1 型。這種易感性使 LC-540 成為病毒學研究的有用模型,特別是在探索病毒與宿主之間的相互作用、病毒致病機制以及開發抗病毒策略方面。

基於其特定特性,LC-540 細胞在包括癌症研究與病毒學在內的一系列研究應用中發揮關鍵作用,有助於深入理解腫瘤形成與病毒感染的機制。

Tissue
睪丸
Disease
腺瘤
Synonyms
  • LC540
  • LC 540

02Classification

Organism
Rat
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Cell type
萊迪格
Tumourigenic
是的,在大鼠身上

04Identifiers

Citation
LC-540(Cytion 型號 500262)
NCBI TaxID
10116
Cellosaurus
CVCL_3536

05Expression and susceptibility

Reverse transcriptase
正向
Products
類固醇激素、雌激素(包括雌二醇等)及雄激素(包括睪固酮等)

06Handling and culture

Culture medium
EMEM(MEM Eagle),含 2 mM L-谷氨醯胺,含 2.2 g/L NaHCO₃,含 EBSS(Cytion 產品編號 820100a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% FBS 和 1% NEAA
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。
Seeding density
1 至 2 × 10⁶ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 24 小時。
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
費雪
Donor age
成人