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MLE-12
Tissue
肺
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 是一種由遠端呼吸道上皮建立的小鼠肺上皮細胞系,其來源為轉基因小鼠,該小鼠在人類表面活性劑蛋白 C(SP-C)啟動子的控制下,表達猿病毒 40(SV40)大腫瘤抗原。 該細胞系的特徵在於能夠維持肺泡Ⅱ型細胞的某些特性,例如表達對肺表面活性劑合成及肺功能至關重要的表面活性劑蛋白 SP-B 和 SP-C。 MLE-12 細胞亦展現出肺泡 II 型細胞的關鍵形態特徵,包括微絨毛和多囊泡體,儘管在後期傳代時會缺乏某些特徵,例如層狀體。

MLE-12 細胞系廣泛用於研究表面活性劑蛋白的調控、分泌,以及肺部對刺激的反應。它會對各種分泌促進劑(如 ATP 和佛波酯)作出反應而分泌磷脂,模擬了 II 型肺泡細胞功能的某些方面。 雖然這種分泌作用在早期傳代時十分旺盛,但在後期傳代中會逐漸減弱,同時受體介導的反應也會發生變化。此模型對於探索呼吸窘迫症候群及表面活性劑缺乏症的潛在機制尤具價值。此外,鑑於該細胞系源自 SV40 驅動的腫瘤形成,亦能為肺癌發生機制提供重要見解。

MLE-12 細胞可用作闡明表面活性劑蛋白質加工途徑,以及測試表面活性劑替代療法策略的工具。由於該細胞系能維持 SP-C 的表達(此為肺泡上皮的特異性標記物),使其成為研究肺部特異性過程與疾病的相關體外模型。

Tissue
肺
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
上皮樣
Cell type
上皮細胞
Tumourigenic
是的,在裸鼠身上

04Identifiers

Citation
MLE-12(Cytion 型號 305314)
NCBI TaxID
10090
Cellosaurus
CVCL_3751

05Expression and susceptibility

Protein expression
表達的基因:肺表面活性蛋白 B、C(SP-B、SP-C)
Viruses
轉化體:猿病毒 40 型(SV40)

06Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
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。
Fluid renewal
每週 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

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
FVB/N-Tg(SFTPC-TAg)5.1Jaw 轉基因小鼠
Donor age
5個月

10Product details

Composition table