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AtT-20
Tissue
脑垂体
Disease
小鼠垂体肿瘤
Synonyms
AtT20, AtT 20, ATT-20

Documents

AtT-20

Cell line · Product number 305161

The AtT-20 cell line is a well-characterized mouse pituitary tumor cell line derived from anterior pituitary cells. These cells originate from a strain of...

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01General information

Description

AtT-20 细胞系是一种特性良好的小鼠垂体瘤细胞系,来源于垂体前叶细胞。这些细胞来源于一种被称为 AtT-20/D16v-F2 的小鼠品系,主要用于研究垂体功能和调节,尤其侧重于促肾上腺皮质激素(ACTH)的合成和分泌。促肾上腺皮质激素对肾上腺功能至关重要,是应激反应和新陈代谢调节的关键因素。

AtT-20 细胞表现出对神经内分泌学和药理学研究具有重要意义的典型特征,如产生和分泌促肾上腺皮质激素的前体分子--原促肾上腺皮质激素(POMC)。这种细胞对促肾上腺皮质激素释放激素(CRH)和其他下丘脑激素有反应,是体外探索下丘脑-垂体-肾上腺(HPA)轴的绝佳模型。此外,AtT-20 细胞的分泌途径明确,可用于研究肽类激素的加工、包装和分泌机制。

在应用方面,AtT-20 细胞已被用于各种研究,包括不同处理条件下的基因表达谱、涉及 cAMP 的细胞内信号通路以及基因修饰对激素分泌的影响。这些细胞在评估针对 HPA 轴成分的潜在候选药物的药理特性方面也很有价值。

Tissue
脑垂体
Disease
小鼠垂体肿瘤
Synonyms
  • AtT20
  • AtT 20
  • ATT-20

02Classification

Organism
Mouse
Growth properties
Suspension
Biosafety level
1

03Characteristics

Morphology
小圆形细胞

04Identifiers

Citation
AtT-20(Cytion 产品编号 305161)
NCBI TaxID
10090
Cellosaurus
CVCL_2300

05Expression and susceptibility

Protein expression
促肾上腺皮质激素(Acth)

06Handling and culture

Culture medium
Ham's F12K 培养基,w:2.0 mM L-谷氨酰胺,w:2.0 mM 丙酮酸钠,w:2.5 g/L NaHCO3(Cytion 文章编号 820608a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 2.5% FBS 和 15% 马血清
Dissociation reagent
Accutase
Subculturing
通过定期添加或更换培养基来维持细胞培养。初始培养时细胞密度应为5×10⁵个细胞/毫升,并保持细胞浓度在3×10⁵至1×10⁶个细胞/毫升范围内以获得最佳生长效果。
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
LAF1