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RAG
Tissue
肾脏
Disease
小鼠肾癌
Synonyms
Rag

Documents

RAG

Cell line · Product number 305190

The RAG cell line is a nonreverting 8-azaguanine-resistant mutant derived from a renal adenocarcinoma of BALB/c mice. This line was developed through alte...

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01General information

Description

RAG 细胞系是一种抗 8-氮杂鸟嘌呤的非逆转突变体,来源于 BALB/c 小鼠的肾腺癌。该细胞系是通过动物到组织培养的交替传代来富集致瘤群体,同时剔除正常的基质成纤维细胞。RAG 细胞的形态从卵球形到上皮样,胞质突起明显,由于缺乏酶,它们对次黄嘌呤-鸟嘌呤磷酸核糖转移酶(HGPRT)依赖性选择方法有抵抗力。这种抗性促进了它们在体细胞杂交实验的生化选择系统中的应用。

由于 RAG 细胞与使用灭活仙台病毒的融合程序兼容,因此在体细胞融合研究中被广泛用作亲本系。与 LM(TK-) 或 WI-38 等其他细胞系融合后,杂交种保留了标记染色体,并表现出代谢缺陷的生化互补性。这些杂交种有助于绘制基因调控元件图谱和研究基因表达,特别是肾脏相关酶,如 ES-2 酯酶。RAG 杂交种为染色体的种间和种内分离以及功能基因组学提供了见解。

除了在杂交研究中的作用外,RAG 细胞还是研究基因表达的表观遗传调控的模型。涉及 RAG 的杂交细胞往往会根据特定染色体的保留或缺失情况,表现出特定遗传性状的消亡和重新表达。这使得 RAG 细胞系成为了解肿瘤细胞基因调控动态和染色体稳定性的重要工具。

Tissue
肾脏
Disease
小鼠肾癌
Synonyms
  • Rag

02Classification

Organism
Mouse
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
Amoeboid

04Identifiers

Citation
RAG(Cytion 产品编号 305190)
NCBI TaxID
10090
Cellosaurus
CVCL_3575

05Expression and susceptibility

Protein expression
肾特异性酯酶-2(ES-2)

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
BALB/c