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12Z
Tissue
子宮內膜、上皮
Disease
子宮內膜異位症
Synonyms
12z, 12-Z, Z12, Z-12, Z12 Eo, EEC12Z

Documents

12Z

Cell line · Product number 305733

The 12Z cell line is an immortalized human endometriotic epithelial cell model derived from peritoneal endometriotic lesions. It was originally establishe...

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  • CoA
1

01General information

Description

12Z 細胞系是一種源自腹膜子宮內膜異位病灶的不朽化人類子宮內膜異位上皮細胞模型。該細胞系最初是透過將 SV40 大 T 抗原轉染至原代子宮內膜異位上皮細胞而建立,從而賦予其延長的增殖能力。 12Z 細胞為細胞角蛋白陽性且 E-鈣黏蛋白陰性,這使其成為具有侵襲性表型的上皮樣細胞群。 研究顯示,這些細胞在體外展現出高度的遷移與侵襲行為,類似於轉移性癌細胞,並表達 N-鈣黏蛋白——這是一種與侵襲性及運動性增強相關的鈣黏蛋白。此分子特徵支持將其用於研究與子宮內膜異位症相關的侵襲機制,並與癌症生物學中的現象相呼應。

在功能方面,12Z 細胞表達參與雌激素和黃體酮訊號傳導、細胞外基質重塑、血管生成、細胞因子產生,以及前列腺素 E2(PGE2)生物合成與訊號傳導的基因。 這些細胞展現出基質金屬蛋白酶 MMP-2 和 MMP-9 的活性升高,這兩種酶對於降解細胞外基質成分及促進組織侵襲至關重要。 此外,12Z 細胞會大量產生 PGE2——這是一種與子宮內膜異位症病理生理學有關的發炎介質。這些特徵,加上其對類固醇激素的反應性,使 12Z 細胞成為剖析子宮內膜異位病灶形成、侵襲及荷爾蒙調控之分子基礎的有效體外模型。

重要的是,近期透過短串聯重複序列(STR)分析進行的品質控制研究,已確認 12Z 細胞的遺傳真實性,從而消除了先前對子宮內膜異位症研究中可能發生交叉污染與誤認的疑慮。 這些細胞,連同其密切相關的 Z11 細胞系,已被提議作為標準模型,用以提升生殖生物學及子宮內膜異位症研究領域的可重複性與可靠性。

Tissue
子宮內膜、上皮
Disease
子宮內膜異位症
Synonyms
  • 12z
  • 12-Z
  • Z12
  • Z-12
  • Z12 Eo
  • EEC12Z

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This cell line contains an SV40 Large T Antigen expression construct delivered via a pcDNA3.1 vector, enabling extended proliferation through p53 and Rb inactivation. The insert is integrated into the human endometriotic cell line 12Z. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮樣
Cell type
上皮細胞
Doubling time
31 小時

04Identifiers

Citation
12Z(Cytion 型號 305733)
NCBI TaxID
9606
Cellosaurus
CVCL_0Q73

05Handling 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)
Seeding density
1–3 × 10⁴ 個細胞/cm²
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.

06Quality 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。

07Donor

Donor age
37年