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LCLC-103H 細胞
Tissue
肺
Disease
大細胞癌
Metastatic site
胸腔積液
Synonyms
LCLC103H, Large Cell Lung Cancer-103H

Documents

LCLC-103H 細胞

Cell line · Product number 300169

The LCLC-103H cell line is derived from a large cell lung carcinoma (LCLC), specifically established from the pleural effusion of an adult male patient wi...

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  • CoA
1

01General information

Description

LCLC-103H 細胞系源自大細胞肺癌(LCLC),具體而言是從一名被診斷為巨細胞型大細胞肺癌的成年男性患者的胸腔積液中建立而來。 該患者先前曾接受過化療和放射治療。此細胞系的特別之處在於其部分表達神經內分泌標記物,這些標記物通常與小細胞肺癌(SCLC)及某些神經內分泌腫瘤相關。 特別是,由單克隆抗體 RNL-1 檢測到的抗原在 LCLC-103H 細胞中呈現局灶性表面表達,這與某些神經內分泌癌所觀察到的現象相似。然而,這種表達並非在所有細胞中均勻分布,表明細胞群體內存在異質性。

文獻中描述 LCLC-103H 為 PAS(過碘酸-席夫)陰性,這使其有別於其他肺癌亞型。該細胞系亦展現顯著的基質形成,這是其組織病理學特徵的重要標誌。 此外,已知該細胞系會過度表達原致癌基因 MYC,該基因在細胞增殖與腫瘤形成過程中扮演關鍵角色。 免疫細胞化學研究顯示,LCLC-103H 並未呈現小細胞肺癌(SCLC)所見的完整神經內分泌分化譜,因其缺乏與其他神經內分泌標記物的反應性,例如由 RNL-2 和 RNL-3 抗體所識別的標記物。 此差異對於區分 LCLC 與小細胞肺癌(SCLC)至關重要,因為後者更具侵襲性,且通常對某些化療藥物展現更高的敏感性。LCLC-103H 獨特的表達模式,使其成為研究大細胞肺癌的分子與免疫學特徵,及其與神經內分泌特徵重疊現象的寶貴模型。

Tissue
肺
Disease
大細胞癌
Metastatic site
胸腔積液
Synonyms
  • LCLC103H
  • Large Cell Lung Cancer-103H

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
多形性
Ploidy status
非整倍體
Doubling time
26 小時

04Identifiers

Citation
LCLC-103H(Cytion 型號 300169)
NCBI TaxID
9606
Cellosaurus
CVCL_1375

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% 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
0.5 至 1 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
細胞將在 24 小時內從冷凍狀態中恢復。
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

Donor age
61年
Ethnicity
高加索人