Skip to content
SK-MEL-5 细胞
Tissue
皮肤
Disease
黑色素瘤
Metastatic site
腋窝淋巴结
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 细胞

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
这是 T. Takahashi 及其同事分离出的大量黑色素瘤细胞系之一。这些品系是检测黑色素瘤患者体内黑色素瘤特异性抗体的靶细胞来源。
Tissue
皮肤
Disease
黑色素瘤
Metastatic site
腋窝淋巴结
Applications
黑色素瘤研究;皮肤癌生物学;黑色素瘤药物敏感性;免疫检查点抑制剂评估;MAPK/ERK通路研究;细胞迁移与侵袭;异种移植模型;自然杀伤细胞和T细胞细胞毒性检测(靶细胞)
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
星状
Cell type
黑色素细胞/黑色素瘤细胞
Tumourigenic
是,在裸鼠体内形成恶性黑色素瘤
Doubling time
约24至36小时

04Identifiers

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

05Expression and susceptibility

Protein expression
P53 阳性
Isoenzymes
PGM1,1-2,PGM3,1,ES-D,1,AK-1,1,GLO-1,1-2,G6PD,B,表型频率产品: 0.0860
Products
黑色素

06Handling and culture

Culture medium
EMEM(MEM Eagle),w:2 mM L-谷氨酰胺,w:2.2 g/L NaHCO3,w:EBSS(Cytion 文章编号 820100a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS 和 1% NEAA
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
Split ratio
1 至 3
Seeding density
1 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 2 至 3 次
Post-thaw recovery
解冻后,将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中,使细胞从冷冻过程中恢复并至少贴壁24小时。
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

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。

Freeze medium
我们使用完全生长培养基(包括 FBS)+10% DMSO 或 CM-1(Cytion 商品目录编号 800100)作为冷冻保存培养基,以获得足够的解冻后存活率,CM-1(Cytion 商品目录编号 800100)含有优化的渗透保护剂和代谢稳定剂,可提高恢复能力并减少冷冻引起的应激。
Storage conditions

如需长期保存,可将样品瓶放入气相液氮中,温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运,并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后,应立即检查容器,并立即将小瓶转移到适当的储藏室。

08Donor

Donor age
24 年
Ethnicity
高加索人