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E11
Tissue
肾脏

Documents

E11

Cell line · Product number 400494

The E11 cell line is a highly specialized murine cell line developed for advanced studies in podocyte function and kidney disease mechanisms. Derived from...

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01General information

Description

E11 细胞系是一种高度特化的小鼠细胞系,用于荚膜细胞功能和肾病机制的高级研究。E11 细胞系来源于表达对温度敏感的 SV40 大 T 抗原变体的转基因小鼠肾小球,在 IFN 诱导的 H-2kb 启动子的调控下运行。这种独特的遗传框架有助于细胞根据环境温度进行有条件增殖,这与 T 抗原的受控表达相一致。

E11 细胞系的一个显著特点是其在大量传代过程中的表型稳定性。经过 40 多次传代后,E11 细胞仍能保持一致的表达和细胞特征,这已被证明是长期研究的宝贵财富,而不会出现许多培养细胞系常见的表型漂移问题。这种稳定性提高了它们在需要细胞行为一致的重复性和扩展性生物实验中的应用。

在蛋白质表达方面,E11 细胞系表现出强大的特征,是进行荚膜特异性研究的必要条件。这种细胞能持续表达荚膜细胞裂隙隔膜结构的重要组成部分--肾素,以及其他多种荚膜细胞特异性蛋白,如 podocin、CD2AP 和 synaptopodin。这种全面的蛋白质表达方式有助于在受控的体外环境中研究荚膜细胞生物学,从而近似模拟体内条件。E11 细胞能形成广泛的细胞-细胞接触,这进一步强调了它们适合用于模拟肾脏滤过屏障功能。

Tissue
肾脏

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This Immorto Mouse podocyte line contains a temperature‑sensitive SV40 T‑antigen construct enabling conditional immortalization. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Cell type
荚膜细胞

04Identifiers

Citation
E11(Cytion 目录编号 400494)
NCBI TaxID
10090
Cellosaurus
CVCL_5737

05Expression and susceptibility

Protein expression
WT1、Lmx1b、肾素、NEPHI、FAT、P-cadherin、CD2AP、ZO-I、podocalyxin、podoplanin、synpo、podocin、TRPC6 和 GAPDH。

06Handling and culture

Culture medium
RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
Incubation atmosphere

33°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
接种T75细胞培养瓶,细胞密度为1×10⁴个细胞/平方厘米,用于增殖过程。将细胞置于33℃/5% CO₂环境中培养,直至培养瓶细胞汇合度达到约75%。
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

Breed / subspecies
(CBA/Ca x C57BL/10)Tg(H2KbtsA58)
Donor age
成人