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B16
Tissue
皮肤
Disease
小鼠黑色素瘤
Synonyms
B-16, B16 melanoma, B16 subline B78, B78

Documents

B16

Cell line · Product number 305154

The B16 cell line is a widely used murine model derived from melanoma tumors in C57BL/6 mice. This line is extensively employed in research due to its abi...

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1

01General information

Description

B16 细胞系是一种广泛使用的小鼠模型,来源于 C57BL/6 小鼠的黑色素瘤肿瘤。该细胞系能形成黑色素瘤,在生长特性和转移潜力方面与人类黑色素瘤非常相似,因此被广泛用于研究。该细胞系有多种亚型,如 B16-F0、B16-F1 和 B16-F10,每种亚型都有不同程度的转移能力;例如,与 B16-F0 相比,B16-F10 的转移能力很强。这些变化使研究人员能够根据他们对肿瘤侵袭性和转移性研究的具体要求选择合适的模型。

B16 细胞对于了解黑色素瘤的分子和细胞进展机制以及测试抗癌疗法非常重要。它们的黑色素生成能力使其在黑色素生成及其调控的研究中尤为有用。此外,B16 细胞系还是疫苗开发和免疫疗法实验的重要工具,有助于深入了解肿瘤与免疫系统的相互作用以及免疫调节药物的功效。这些细胞对各种体内和体外环境的适应性突显了它们在旨在治疗和预防黑色素瘤的转化和临床前研究中的重要性。

Tissue
皮肤
Disease
小鼠黑色素瘤
Synonyms
  • B-16
  • B16 melanoma
  • B16 subline B78
  • B78

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
纺锤形细胞和上皮样细胞的混合物
Tumourigenic
是

04Identifiers

Citation
B16(Cytion 产品编号 305154)
NCBI TaxID
10090
Cellosaurus
CVCL_F936

05Expression and susceptibility

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。
Fluid renewal
每周 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

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

Breed / subspecies
C57BL/6