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mIMCD-3
Tissue
腎臟,內髓質集合管
Disease
正常
Synonyms
mIMCD3, IMCD-3, IMCD3, mouse Inner Medullary Collecting Duct-3

Documents

mIMCD-3

Cell line · Product number 305382

The mIMCD-3 cell line is derived from the murine inner medullary collecting duct and is widely used as a model for studying renal epithelial cell biology,...

US$540.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

mIMCD-3 細胞系源自小鼠內髓質集合管,廣泛用作研究腎上皮細胞生物學、腎臟發育及病理生理學的模型。 此永生化細胞系展現出集合管典型的特徵,包括能在三維培養系統中形成管狀結構,以及表達緊密連接蛋白、水通道蛋白和上皮標記物。 mIMCD-3 細胞對於探索離子轉運、水分調節以及細胞對機械應力的反應機制尤為寶貴,使其成為腎臟學研究中不可或缺的工具。

mIMCD-3 細胞已被應用於多囊腎(PKD)的研究,以釐清囊腫形成的分子基礎。 特別是,該細胞系經過基因改造,用於探究多囊蛋白(PKD1 和 PKD2)的作用,這些蛋白質是常染色體顯性多囊腎(ADPKD)中的關鍵因子。 舉例來說,缺乏 PKD1 的 mIMCD-3 細胞在受到機械應力時會出現異常的腎小管擴張,這與 PKD 患者所觀察到的囊腫形成現象相仿。此外,涉及 mIMCD-3 細胞的實驗已證實,初級纖毛在介導 cAMP 訊號傳導與機械轉導方面扮演重要角色,而這兩者皆是腎上皮功能及疾病病理學中的關鍵過程。

該細胞系亦被應用於藥物發現與測試,特別是在腎毒性及腎臟特異性治療的領域。mIMCD-3 細胞為研究對順鉑等化療藥物的反應提供了平台,此類藥物可能在腎細胞中誘發氧化壓力與細胞凋亡。 此外,mIMCD-3 細胞在培養條件下展現的強健生長特性與可重複性,使其成為研究腎臟生物學各個面向,以及推動腎臟疾病治療方法發展的靈活模型。

Tissue
腎臟,內髓質集合管
Disease
正常
Synonyms
  • mIMCD3
  • IMCD-3
  • IMCD3
  • mouse Inner Medullary Collecting Duct-3

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine inner medullary collecting duct cell line (mIMCD-3) contains an SV40 T-Antigen transgene derived from a transgenic mouse model, supporting immortalization. The construct is stably present in primary kidney-derived cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
上皮樣

04Identifiers

Citation
mIMCD-3(Cytion 型號 305382)
NCBI TaxID
10090
Cellosaurus
CVCL_0429

05Expression and susceptibility

Viruses
轉化體:猿病毒 40 型(SV40)

06Handling and culture

Culture medium
DMEM:Ham's F12(1:1),重量比:3.1 g/L 葡萄糖,重量比:2.5 mM L-谷氨酰胺,重量比:15 mM HEPES, w: 0.5 mM 丙酮酸鈉,w: 1.2 g/L NaHCO₃(Cytion 產品編號 820400a)
Incubation atmosphere

37°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
1 至 3 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 2 至 5 × 10⁴ 個細胞/cm² 的密度接種至培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 24 小時。
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
Tg(SV40E)Bri/7 轉基因
Donor age
成人