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FS-C3H
Tissue
皮肤
Disease
纤维肉瘤

Documents

FS-C3H

Cell line · Product number 400418

The FS-C3H cell line, derived from the C3H/HeJ mouse strain, plays a pivotal role in studying host responses to endotoxins, particularly in the context of...

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01General information

Description

FS-C3H细胞系来源于C3H/HeJ小鼠品系,在研究宿主对内毒素的反应,尤其是癌症研究方面发挥着关键作用。该品系对细菌内毒素的主要成分脂多糖(LPS)特异性不敏感,因而对内毒素具有显著的抵抗力。这一特性使 FS-C3H 成为剖析免疫反应调节所涉及的生化和遗传途径的宝贵模型。研究人员广泛使用这种细胞系来研究 B 淋巴细胞和巨噬细胞的动态,重点研究它们对 LPS 的独特无反应性,这与典型的免疫细胞对此类刺激的反应形成鲜明对比。

FS-C3H 细胞对 LPS 无反应是因为缺乏或改变了负责 LPS 信号转导的关键受体。研究表明,尽管这些细胞对 LPS 没有反应,但它们可以通过蛋白激酶 C(PKC)和酪氨酸激酶信号机制等替代途径被激活,这些途径与 LPS 反应细胞激活的途径相似。这些激酶在信号通路中的相互作用和调节作用凸显了复杂的细胞内机制,表明 PKC 和酪氨酸激酶通路可以弥补 LPS 信号的缺陷。这一观察结果为探索酪氨酸激酶调节的磷酸化如何影响这些小鼠的整体细胞反应开辟了途径。

继续对 FS-C3H 细胞进行研究对于了解它们对 LPS 反应低下的分子基础至关重要,这可能与 Lpsn 基因的遗传缺陷有关。通过深入研究这些细胞与 LPS 反应者相比的磷酸化特征,科学家们旨在揭示导致基因激活和增殖反应改变的特定分子缺陷。通过分离和鉴定导致 LPS 相互作用的基因产物,可以深入了解免疫系统的功能障碍,并为治疗相关免疫和炎症性疾病的新型治疗方法铺平道路。

Tissue
皮肤
Disease
纤维肉瘤

02Classification

Organism
Mouse
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
FS-C3H(Cytion 目录编号 400418)
NCBI TaxID
10090
Cellosaurus
CVCL_5755

04Handling 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
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.

05Quality 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。

06Donor

Breed / subspecies
C3H