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SK-N-MC
Tissue
神經外胚層
Disease
阿斯金氏腫瘤
Metastatic site
眶上區
Synonyms
SKNMC, SK-NM-C, SK-NMC

Documents

SK-N-MC

Cell line · Product number 300340

This cell line has been established by J.L. Biedler in 1971. It has moderate dopamine-beta-hydroxylase activity as well as formaldehyde induced fluorescence ...

US$395.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description
該細胞系由 J.L. Biedler 於 1971 年建立。其具有中等程度的多巴胺-β-羥化酶活性,並會因甲醛誘導而產生螢光,這表明細胞內存在兒茶酚胺。
Tissue
神經外胚層
Disease
阿斯金氏腫瘤
Metastatic site
眶上區
Synonyms
  • SKNMC
  • SK-NM-C
  • SK-NMC

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
成纖維細胞樣
Tumourigenic
是的,在裸鼠以及倉鼠頰囊中
Karyotype
低二倍體至假二倍體。異常現象包括雙分鐘染色體、斷裂、大型亞中著絲點染色體、端著絲點染色體及小型端著絲點染色體標記(原始來源)。 (P32) 低二倍體至超二倍體,以及三倍體至低四倍體,異常現象包括雙著絲點染色體、斷裂、雙分鐘染色體(DM)、大型亞端粒染色體及小型端粒染色體。
Doubling time
32 小時

04Identifiers

Citation
SK-N-MC(Cytion 型號 300340)
NCBI TaxID
9606
Cellosaurus
CVCL_0530

05Expression and susceptibility

Antigen expression
O型血,Rh陽性
Isoenzymes
Me-2,2;PGM3,1-2;PGM1,1;ES-D,2;AK-1,1;GLO-1,1-2;G6PD,B

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
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 至 2 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 24 小時。
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 50% 基礎培養基 + 40% 胎牛血清 (FBS) + 10% 二甲基亞砜 (DMSO),或 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,可提高細胞存活率並減輕冷凍引起的應激。
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
12年
Ethnicity
高加索人