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Tissue
小肠,回肠
Disease
正常
Synonyms
IEC 18, IEC18, Intestinal Epithelioid Cell line No. 18

Documents

IEC-18

Cell line · Product number 305302

US$395.00excl. tax and shipping
Made to order
1

01General information

Description

IEC-18 细胞系是一种非转化上皮细胞系,来源于大鼠小肠的隐窝细胞。这些细胞已被证明能有效模拟小肠上皮细胞的生理特性,尤其是氯离子(Cl-)转运特性。IEC-18 细胞中的氯离子通道表现出不同类型的传导,可对细胞肿胀、细胞内钙(Ca²⁺)增加和环磷酸腺苷(cAMP)升高等各种刺激做出反应。例如,IEC-18 细胞中膨胀激活的 Cl- 电流具有外向整流和电压无关的特点。此外,IEC-18 细胞还表达囊性纤维化跨膜传导调节器(CFTR)通道,这体现在细胞中存在 cAMP 激活的 Cl- 传导,格列本脲和 5-硝基-2-(3-苯基丙基氨基)苯甲酸(NPPB)可抑制这种传导,但 DIDS 不会影响这种传导。

IEC-18 细胞还被用于探索脱落诱导应激(即 anoikis)下的细胞存活机制。研究表明,前列腺素 E2(PGE2)可通过 cAMP 介导的信号通路促进脱落 IEC-18 细胞的存活和聚集。这种保护作用与腺苷酸环化酶和蛋白激酶 A(PKA)的激活有关,即使在悬浮状态下也能增强细胞的粘附性和活力。这些发现对于了解炎症相关过程以及肠道组织中潜在的致癌因素具有重要意义。

此外,IEC-18 单层细胞还被用来研究各种分子通过肠道屏障的转运。与 Caco-2 细胞系相比,IEC-18 细胞在结构上与小肠隐窝细胞相似,因此为被动跨细胞和旁细胞转运提供了更准确的模型。与具有显著主动转运能力的 Caco-2 细胞不同,IEC-18 细胞显示出最小的载体介导转运能力,因此更适合用于分析亲水性大分子的被动渗透性。

Tissue
小肠,回肠
Disease
正常
Synonyms
  • IEC 18
  • IEC18
  • Intestinal Epithelioid Cell line No. 18

02Classification

Organism
Rat
Sex
Unspecified
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样
Cell type
上皮细胞

04Identifiers

Citation
IEC-18(Cytion 目录编号 305302)
NCBI TaxID
10116
Cellosaurus
CVCL_0342

05Handling 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 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.

06Quality 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。

07Donor

Breed / subspecies
Charles River Sprague Dawley (CD(SD))
Donor age
18-24 天

09Product details

Composition table