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H4-II-E-C3
Tissue
肝臟
Disease
肝細胞癌
Synonyms
H4IIEC3, H4-IIE-C3, H4-II-EC3, H-II-E-C3

Documents

H4-II-E-C3

Cell line · Product number 305336

The H4-II-E-C3 cell line is a well-established rat hepatoma cell line derived from liver carcinoma. This line is widely used in research focused on liver-...

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Lot
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  • CoA
1

01General information

Description

H4-II-E-C3 細胞系是一種源自肝癌的、已確立的大鼠肝癌細胞系。該細胞系廣泛應用於針對肝臟特異性功能及肝細胞內代謝過程的研究。 H4-II-E-C3 細胞的一個顯著特徵是其高親和力谷氨酸轉運蛋白 GLT-1A 的高表達。此特徵使其有別於正常大鼠肝細胞,後者並未表現出顯著水平的此種轉運蛋白。 H4-II-E-C3 細胞中 GLT-1A 的表達,使研究人員能有效探究谷氨酸的代謝與轉運機制,從而深入了解肝細胞的轉化過程及腫瘤細胞的代謝狀況。

此外,H4-II-E-C3 細胞亦被用於研究醛類代謝,特別是線粒體第 2 類醛類脫氫酶(ALDH2)的活性。該酶對於代謝乙醇氧化過程中產生的乙醛至關重要,因此該細胞系非常適合用於研究酒精代謝及其對細胞的影響。 在這些細胞中採用「乙醛夾」技術,無需分離線粒體即可精確測量 ALDH2 活性,從而促進了酶動力學研究,以及各類抑制劑對醛類解毒作用的影響研究。

Tissue
肝臟
Disease
肝細胞癌
Synonyms
  • H4IIEC3
  • H4-IIE-C3
  • H4-II-EC3
  • H-II-E-C3

02Classification

Organism
Rat
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This rat hepatocellular carcinoma cell line (H4-II-E-C3) contains undefined modifications classified as GMO, stably present in the transformed hepatic cell line. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Tumourigenic
是的,在 AxC 大鼠中

04Identifiers

Citation
H4-II-E-C3(Cytion 型號 305336)
NCBI TaxID
10116
Cellosaurus
CVCL_0285

05Expression and susceptibility

Protein expression
基因表達:酪胺酸氨基轉移酶、白蛋白、轉鐵蛋白、凝血酶原

06Handling 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。
Seeding density
1-3×10⁴/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.

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
AxC
Donor age
未指定