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NCI-H209 細胞
Tissue
肺
Disease
小細胞癌
Metastatic site
骨髓
Synonyms
H209, H-209, NCIH209

Documents

NCI-H209 細胞

Cell line · Product number 300183

The NCI-H209 cell line was derived by A.F. Gazdar and associates in 1979 from the bone marrow of a patient with small cell cancer of the lung. The bone marro...

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1

01General information

Description
NCI-H209 細胞系由 A.F. Gazdar 及其同事於 1979 年從一名小細胞肺癌患者的骨髓中分離獲得。該骨髓檢體是在治療前採集的。 該細胞系為典型的微小細胞肺癌(SCLC)細胞系,其表現出四種生化標記物(神經特異性烯醇化酶、腦型肌酸激酶同工酶、L-多巴脫羧酶及邦貝辛樣免疫反應性)的升高水平。C-myc DNA 序列未被擴增。 未檢測到明顯的DNA結構異常。此細胞系在懸浮培養中會形成大型聚集体生長。僅聚集体具有存活能力,但無法測量出具意義的存活率百分比。培養基中通常含有大量細胞碎片。 這些細胞表達一種未經磷酸化處理的異常 RB1 形式,顯然是由於第 706 編碼子處發生單點突變(Cys→Phe)所致。
Tissue
肺
Disease
小細胞癌
Metastatic site
骨髓
Synonyms
  • H209
  • H-209
  • NCIH209

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
Biosafety level
1

03Characteristics

Morphology
上皮樣
Tumourigenic
是的,會在裸鼠體內形成具有典型小細胞肺癌(SCLC)組織學特徵的可移植腫瘤

04Identifiers

Citation
NCI-H209(Cytion 型號 300183)
NCBI TaxID
9606
Cellosaurus
CVCL_1525

05Expression and susceptibility

Protein expression
P53 陰性
Isoenzymes
G6PD、B、PGM1、1-2、PGM3、1、ES-D、1、Me-2、0、AK-1、1、GLO-1、1-2,表型頻率積 = 0.0624
Products
與正常肺組織相比,該細胞系所產生的 p53 mRNA 量處於正常範圍內。

06Handling 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)
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 × 105 個細胞/毫升
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

Donor age
55年
Ethnicity
高加索人