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MLE-12
Tissue
肺部
Disease
正常
Synonyms
MLE 12, MLE12, Murine Lung Epithelial-12

Documents

MLE-12

Cell line · Product number 305314

US$550.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
  • CoA
1

01General information

Description

MLE-12 是利用转基因小鼠从远端呼吸道上皮建立的小鼠肺上皮细胞系,它在人表面活性物质蛋白 C (SP-C) 启动子的控制下表达猿病毒 40 (SV40) 大肿瘤抗原。这种细胞系的特点是能保持肺泡 II 型细胞的某些特性,如表达对肺表面活性物质合成和肺功能至关重要的表面活性物质蛋白 SP-B 和 SP-C。MLE-12细胞还显示出肺泡II型细胞的主要形态特征,包括微绒毛和多囊体,但它们在后期传代中缺乏片状体等一些特征。

MLE-12 细胞系被广泛用于研究表面活性物质蛋白的调节、分泌和肺对刺激的反应。它能分泌磷脂以应对 ATP 和植物醇酯等各种分泌物,模拟 II 型肺泡细胞功能的各个方面。这种分泌在早期很旺盛,但在后期随着受体介导的反应的变化而减少。这种模型对于探索呼吸窘迫综合征和表面活性物质缺乏症的机制特别有价值。此外,由于该细胞系来源于 SV40 驱动的肿瘤发生,它还能让人们深入了解肺癌的发生。

MLE-12 细胞是阐明表面活性物质蛋白加工途径和测试表面活性物质替代治疗策略的工具。肺泡上皮特异性标志物 SP-C 的表达保持不变,使其成为研究肺部特异性过程和疾病的相关体外模型。

Tissue
肺部
Disease
正常
Synonyms
  • MLE 12
  • MLE12
  • Murine Lung Epithelial-12

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine lung epithelial cell line (MLE-12) contains an SV40 T-Antigen construct introduced via transfection, supporting immortalization of primary lung epithelial cells. The insert is stably integrated. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮样
Cell type
上皮细胞
Tumourigenic
是,在裸鼠身上

04Identifiers

Citation
MLE-12(Cytion 目录编号 305314)
NCBI TaxID
10090
Cellosaurus
CVCL_3751

05Expression and susceptibility

Protein expression
表达的基因:肺表面活性蛋白 B、C(SP-B、SP-C)
Viruses
转化体: Simian virus 40 (SV40)

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 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
FVB/N-Tg(SFTPC-TAg)5.1Jaw 转基因
Donor age
5 个月

10Product details

Composition table