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C127I
Tissue
乳房、乳腺
Disease
癌症
Synonyms
C 1271, C-127I, C-127 I, CNC 127I

Documents

C127I

Cell line · Product number 400134

The C127I cell line is a murine mammary gland epithelial cell line, commonly used in biomedical research for its ability to synthesize and secrete recombi...

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01General information

Description

C127I 细胞系是一种小鼠乳腺上皮细胞系，因其具有合成和分泌重组蛋白的能力而常用于生物医学研究。这些细胞来源于 BALB/c 小鼠的乳腺，其上皮形态以及对激素和其他生长因子的反应能力尤为突出。C127I 细胞系在研究基因表达、与癌症发展相关的信号转导途径以及生产用于基因治疗的病毒载体方面发挥了重要作用。

C127I 细胞系的主要特点之一是易于转染，是生产重组蛋白和进行基因编辑研究的重要工具。它支持各种小鼠逆转录病毒的复制，有利于生产表达理想基因的稳定重组品系。这一特性使 C127I 细胞在分子生物学和遗传学领域特别有用，经常被用来在受控环境中探索基因过表达或基因敲除的效果。

Tissue
乳房、乳腺
Disease
癌症
Applications
用于牛乳头状瘤病毒 DNA 质粒转化的转染宿主。肉瘤病毒诱导灶的可视化。牛乳头瘤病毒的体外定量检测。
Synonyms
  • C 1271
  • C-127I
  • C-127 I
  • CNC 127I

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine breast carcinoma cell line (C127I) contains recombinant viral sequences encoding T7 RNA polymerase and CFTR delivered via infection with engineered viruses, functioning as a transfection host. The construct is stably integrated into C127 cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮样

04Identifiers

Citation
C127I（Cytion 目录编号 400134）
NCBI TaxID
10090
Cellosaurus
CVCL_3882

05Expression and susceptibility

Viruses
外胚叶病毒（鼠痘）阴性。
Virus susceptibility
牛乳头瘤病毒
Reverse transcriptase
阴性（在上清液中测定）

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。
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

Breed / subspecies
RIII