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E11
Tissue
腎臟

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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1

01General information

Description

E11 細胞系是一種高度專一化的小鼠細胞系,專為足細胞功能及腎臟疾病機制之深入研究而開發。 E11 細胞源自經基因工程改造、可表達 SV40 大 T 抗原溫度敏感變異體的轉基因小鼠腎小球,其運作受 IFN-γ 誘導型 H-2kb 啟動子所調控。 這種獨特的遺傳架構有助於細胞根據環境溫度進行有條件的增殖,這與 T 抗原的可控表達相吻合。

E11 細胞系的一大特色在於,即使經過大量傳代,其表型仍能保持穩定。 E11 細胞在超過 40 次傳代過程中仍能維持一致的表達與細胞特性,對於長期研究而言極具價值,且不會出現許多培養細胞系常見的表型漂移問題。這種穩定性使其更適合用於需要細胞行為一致的重複性及長期生物實驗。

就蛋白質表達而言,E11 細胞系展現出強健的表達特徵,這對於足細胞特異性研究至關重要。這些細胞能持續表達足細胞裂隙隔膜結構的關鍵成分——腎素(nephrin),同時也表達多種其他足細胞特異性蛋白質,例如足細胞蛋白(podocin)、CD2AP 及突觸足蛋白(synaptopodin)。 這種全面的蛋白質表達特性,有助於在受控的體外環境中研究足細胞生物學,並能密切模擬體內條件。E11 細胞形成廣泛細胞間接觸的能力,進一步凸顯其作為腎臟過濾屏障功能模型的適用性。

Tissue
腎臟

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
足細胞

04Identifiers

Citation
E11(Cytion 型號 400494)
NCBI TaxID
10090
Cellosaurus
CVCL_5737

05Expression and susceptibility

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

06Handling and culture

Culture medium
RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
Incubation atmosphere

33°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% 的胎牛血清(FBS)
Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
Seeding density
將 T75 細胞培養瓶接種 1×10⁴ 個細胞/cm²,以進行增殖培養。將細胞置於 33 攝氏度/5% CO₂ 的環境中,直至培養瓶內細胞覆蓋率約達 75%。
Fluid renewal
每週 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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

08Donor

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