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LMH
Tissue
肝臟
Disease
肝細胞癌
Synonyms
Leghorn Male Hepatoma cell line

Documents

LMH

Cell line · Product number 601411

LMH cells, derived from a Leghorn male hepatoma, are a versatile cell line widely used in biological research. Tomoyuki Kitagawa established them in 1981 ...

US$550.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

源自萊格霍恩公雞肝癌的 LMH 細胞,是一種廣泛應用於生物研究的多功能細胞系。北川智之於 1981 年在日本東京癌症研究所成功建立此細胞系。這些細胞具有上皮表型,對於研究家禽胃腸道中的宿主-病原體相互作用特別有用。

LMH 細胞為貼壁細胞,呈現樹突狀形態。它們表達葡萄糖-6-磷酸酶,並具有微弱的管狀 ATP 酶活性。這些細胞具有三倍體核型及六條標記染色體,展現出鮮明的遺傳特徵。
值得注意的是,研究顯示當轉染病毒構建體時,LMH細胞能有效支持鴨乙型肝炎病毒(DHBV)的DNA合成。這使它們成為病毒學研究中不可或缺的工具,特別是在家禽相關病毒感染的研究領域。
LMH 細胞的建立過程,是透過長期以二乙基亞硝胺處理,在萊格霍恩雞的肝臟中誘導出腫瘤結節。這些細胞亦經過化學轉化,使其能夠在培養中實現永生化並持續增殖。

就致瘤性而言,LMH 細胞具有在無胸腺裸鼠體內形成腫瘤的能力。此特性使其成為研究肝細胞癌的重要模型。 LMH 細胞會表達雌激素受體,並可被誘導表達肝臟特異性載脂蛋白 II(apoII)基因。這表明它們參與了雌激素訊號傳導途徑及脂質代謝。要培養 LMH 細胞,必須先在組織培養容器上預塗膠原蛋白,以確保細胞能正常黏附與生長。

Tissue
肝臟
Disease
肝細胞癌
Applications
該細胞系適用於轉染研究。
Synonyms
  • Leghorn Male Hepatoma cell line

02Classification

Organism
Chicken
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮樣、樹突樣。
Tumourigenic
LMH 細胞會在無胸腺小鼠體內形成腫瘤。
Karyotype
三倍體,模數 = 116,六條標記染色體

04Identifiers

Citation
LMH(Cytion 型號 601411)
NCBI TaxID
9031
Cellosaurus
CVCL_2580

05Expression and susceptibility

Receptors expressed
雌激素(低水平表達)。
Products
葡萄糖-6-磷酸酶,管腔型 ATP 酶活性(微弱)

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
LMH 細胞在預先塗佈膠原蛋白的組織培養容器上附著效果較佳。移除培養基,並使用不含鈣和鎂的 PBS 沖洗附著的細胞(T25 細胞培養瓶需 3-5 毫升 PBS,T75 細胞培養瓶需 5-10 毫升)。 加入 Accutase(T25 培養瓶每瓶 1-2 毫升,T75 培養瓶每瓶 2.5 毫升),確保細胞片完全被覆蓋。於室溫下孵育 8-10 分鐘。 小心用培養基(10 毫升)將細胞重新懸浮,以 300 g 離心 3 分鐘,再將細胞懸浮於新鮮培養基中,並分裝至裝有新鮮培養基的新培養瓶中。
Seeding density
1 至 3 × 10⁴ 個細胞/cm²
Fluid renewal
每兩天
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
萊格霍恩
Donor age
16個月