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NCI-H82
Tissue
肺
Disease
肺小細胞癌
Metastatic site
胸腔積液
Synonyms
NCI-H-82, H82, H-82, NCI H82, NCIH82, H82sclc

Documents

NCI-H82

Cell line · Product number 300442

The NCI-H82 cell line was derived by A.F. Gazdar and associates in 1978 from the pleural fluid of a patient with small cell cancer of the lung. The morpholog...

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01General information

Description
NCI-H82 細胞系由 A.F. Gazdar 及其同事於 1978 年從一名小細胞肺癌患者的胸腔積液中分離得到。原始腫瘤的形態並無小細胞肺癌(SCLC)的典型特徵。 該細胞系是小細胞肺癌(SCLC)的一種生化與形態學變異型,會表達神經特異性烯醇化酶及肌酸激酶的腦同工酶。其 L-多巴脫羧酶或邦貝辛的含量均未達可檢測水平。這些細胞會產生大小異常的 p53 mRNA(3.7 kb)。 c-myc DNA 序列擴增約 25 倍,且相較於正常細胞,c-myc RNA 量增加 24 倍。據報告,這些細胞表達具有功能的 ANP 受體,但使用 ANP 治療並未改變其生長模式。這些細胞對神經絲蛋白和維曼汀呈陽性染色。 可檢測到 v-fes、v-fms、Ha-ras、Ki-ras、N-ras 及 c-raf 1 mRNA 的表達。
Tissue
肺
Disease
肺小細胞癌
Metastatic site
胸腔積液
Synonyms
  • NCI-H-82
  • H82
  • H-82
  • NCI H82
  • NCIH82
  • H82sclc

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非典型小細胞肺癌(SCLC)組織學特徵的可移植腫瘤
Karyotype
這是一種近三倍體的人類細胞系。其模態染色體數為 58,佔 44%;多倍體比例為 3%。每個細胞均含有兩份正常的 X 染色體。在 Q 帶染色製備物中未檢測到 Y 染色體。

04Identifiers

Citation
NCI-H82(Cytion 型號 300442)
NCBI TaxID
9606
Cellosaurus
CVCL_1591

05Expression and susceptibility

Receptors expressed
胰島素樣生長因子 II 受體(IGF II)、心房鈉尿肽(ANP)
Protein expression
P53 陽性
Isoenzymes
G6PD、B、PGM1、1-2、PGM3、1-2、ES-D、1、Me-2、1、AK-1、1、GLO-1、1,表型頻率積 = 0.0082

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
透過定期添加或更換培養基來維持培養。以 5 × 10⁵ 細胞/毫升的密度啟動培養,並將細胞濃度維持在 3 × 10⁵ 至 1 × 10⁶ 細胞/毫升的範圍內,以達到最佳生長效果。
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
41年
Ethnicity
高加索人