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SK-MEL-5 細胞
Tissue
皮膚
Disease
黑色素瘤
Metastatic site
腋窩淋巴結
Synonyms
SK-Mel-5, SK MEL 5, SK.MEL.5, SK-MEL5, SKMel-5, SKMEL-5, SKMEL5, SKMel5, SKmel5, AA-Mel

Documents

SK-MEL-5 細胞

Cell line · Product number 300157

This is one of a very extensive series of melanoma lines which have been isolated by T. Takahashi and associates. The lines served as source of target cells ...

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

01General information

Description
這是由 T. Takahashi 及其同事所分離出的、極為龐大的黑色素瘤細胞系系列之一。這些細胞系用作檢測黑色素瘤患者體內黑色素瘤特異性抗體的靶細胞來源。
Tissue
皮膚
Disease
黑色素瘤
Metastatic site
腋窩淋巴結
Applications
黑色素瘤研究;皮膚癌生物學;黑色素瘤藥物敏感性;檢查點抑制劑評估;MAPK/ERK 路徑研究;細胞遷移與侵襲;異種移植模型;自然殺傷細胞與 T 細胞細胞毒性檢測(標靶細胞)
Synonyms
  • SK-Mel-5
  • SK MEL 5
  • SK.MEL.5
  • SK-MEL5
  • SKMel-5
  • SKMEL-5
  • SKMEL5
  • SKMel5
  • SKmel5
  • AA-Mel

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
Non-GMO
GMO status details
No genetic modification; wildtype melanoma cell line isolated by T. Takahashi et al.
Biosafety level
1

03Characteristics

Morphology
星狀
Cell type
黑色素細胞/黑色素瘤細胞
Tumourigenic
是的,在裸鼠身上會形成惡性黑色素瘤
Doubling time
約 24 至 36 小時

04Identifiers

Citation
SK-MEL-5(Cytion 型號 300157)
NCBI TaxID
9606
Cellosaurus
CVCL_0527

05Expression and susceptibility

Protein expression
P53 陽性
Isoenzymes
PGM1,1-2,PGM3,1,ES-D,1,AK-1,1,GLO-1,1-2,G6PD,B,表型頻率乘積:0.0860
Products
黑色素

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。
Split ratio
1 至 3
Seeding density
1 × 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
24年
Ethnicity
高加索人