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266-6
Tissue
胰腺
Disease
小鼠胰腺腺泡肿瘤

Documents

266-6

Cell line · Product number 305363

The 266-6 cell line is a mouse pancreatic acinar cell model that has been extensively used in studies investigating pancreatic physiology and pathophysiol...

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01General information

Description

266-6细胞系是一种小鼠胰腺腺泡细胞模型,已被广泛应用于研究胰腺生理学和病理生理学,特别是与胰腺炎和营养物质代谢相关的研究。该细胞系源自小鼠胰腺腺泡细胞,这类细胞专门负责消化酶的合成与分泌。

对 266-6 细胞的研究表明,它们在探索细胞死亡机制方面具有重要价值,其中包括铁死亡——一种由脂质过氧化驱动的受调控坏死。 特别是,研究表明,当 266-6 细胞接触胰蛋白酶、蓝肽或酒精等物质时,会对铁死亡产生敏感性。 这表明,266-6细胞是研究胰腺损伤相关分子通路(特别是在急性或慢性胰腺炎等病症中)的宝贵模型。

此外,266-6细胞还被用于研究营养物质转运机制,例如硫胺素(维生素B1)及其磷酸化形式——硫胺素焦磷酸(TPP)的摄取。研究表明,长期接触酒精或促炎细胞因子等因素会损害这些细胞对硫胺素的摄取。 这种功能障碍发生在硫胺素转运蛋白THTR-1和THTR-2以及线粒体TPP转运蛋白(MTPPT)的转录和蛋白质表达两个层面,从而影响胰腺细胞的新陈代谢,并导致线粒体功能障碍。

Tissue
胰腺
Disease
小鼠胰腺腺泡肿瘤

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This mouse pancreatic acinar cell line contains an SV40 large T antigen expression cassette under control of the elastase-1 promoter, driving immortalization of exocrine pancreatic cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
上皮样
Cell type
胰腺腺泡细胞

04Identifiers

Citation
266-6(Cytion 产品编号 305363)
NCBI TaxID
10090
Cellosaurus
CVCL_3481

05Expression and susceptibility

Receptors expressed
胆碱能乙酰胆碱受体

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
1至3×10⁴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.

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
成人