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HS-695T 细胞
Tissue
皮肤
Disease
黑色素瘤
Metastatic site
淋巴结
Synonyms
Hs 695.T, Hs-695-T, Hs 695T, HS 695T, Hs695T, HS695T, Hs695

Documents

HS-695T 细胞

Cell line · Product number 300211

The HS-695T cell line is derived from human melanoma, a type of skin cancer characterized by the malignant transformation of melanocytes. These cells were...

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01General information

Description

HS-695T 细胞系来源于人类黑色素瘤,这是一种以黑色素细胞恶性转化为特征的皮肤癌。这些细胞最初来自一名成年患者,后来被广泛用于黑色素瘤生物学、肿瘤发生和癌症转移方面的研究。HS-695T 细胞系具有黑色素瘤的主要特征,包括在移植到免疫功能低下的小鼠体内后能迅速增殖并形成肿瘤。这种细胞系保留了原始肿瘤的许多分子和遗传特征,因此是研究黑色素瘤进展内在机制和测试潜在治疗药物的宝贵模型。

HS-695T 细胞表达多种黑色素瘤相关标记物,包括 Melan-A、酪氨酸酶和 HMB-45,这些标记物通常用于识别和研究黑色素细胞肿瘤。据了解,这些细胞还存在 BRAF 和 NRAS 等基因突变,这些基因突变在黑色素瘤中经常出现,是驱动肿瘤生长和存活的致癌信号通路。研究人员利用 HS-695T 细胞系探索 BRAF 和 MEK 抑制剂等靶向疗法的效果,并研究这些疗法的抗药性发展。总之,HS-695T 细胞系是黑色素瘤研究的重要工具,它有助于发现新的治疗策略,提高我们对这种侵袭性癌症的认识。

Tissue
皮肤
Disease
黑色素瘤
Metastatic site
淋巴结
Synonyms
  • Hs 695.T
  • Hs-695-T
  • Hs 695T
  • HS 695T
  • Hs695T
  • HS695T
  • Hs695

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样
Tumourigenic
是,在免疫抑制小鼠中
Karyotype
(P19-40) 模式 = 52,存在 Y 染色体

04Identifiers

Citation
HS-695T(Cytion 目录号 300211)
NCBI TaxID
9606
Cellosaurus
CVCL_0851

05Expression and susceptibility

Protein expression
P53 阳性
Isoenzymes
G6PD,B,PGM1,1,PGM3,1,ES-D,1,Me-2,0,AK-1,1,GLO-1,1,表型频率产品: 0.0427
Mutational profile
BRAF V600Emut

06Handling and culture

Culture medium
DMEM,w:4.5 克/升葡萄糖,w:4 毫摩尔 L-谷氨酰胺,w:3.7 克/升 NaHCO3,w:1.0 毫摩尔丙酮酸钠(Cytion 编号 820300a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
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。
Seeding density
2 × 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
26 年
Ethnicity
高加索人