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GIST-T1
Tissue
转移性
Disease
胃肠道间质瘤
Metastatic site
胸腔积液
Synonyms
GIST-T-1, GISTT1, T1

Documents

GIST-T1

Cell line · Product number 305777

The GIST-T1 cell line is a well-established human gastrointestinal stromal tumor (GIST) model derived from a metastatic pleural lesion secondary to a prim...

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1

01General information

Description

GIST-T1细胞系是一种成熟的人胃肠道间质瘤(GIST)模型,源自一名日本成年女性原发性胃GIST引起的转移性胸膜病变。 免疫组织化学分析证实,该细胞系对 GIST 的两个标志性标志物 c-KIT (CD117) 和 CD34 呈强阳性,而对肌动蛋白、S-100 和 α-平滑肌肌动蛋白呈阴性,证实了其非肌肉和非神经起源。 细胞遗传学研究显示该细胞系具有低二倍体核型,伴有复杂的染色体异常,包括环状染色体和若干不平衡易位。 比较基因组杂交(CGH)和荧光原位杂交(FISH)分析显示,在 3q26.1-27、5p12-15.1 和 7q21.3-36 区域存在高水平扩增,这些区域通常与 GIST 中的致癌基因扩增有关。

GIST-T1 携带 *KIT* 基因第 11 外显子中一个具有临床意义的 57 核苷酸同框缺失(V570-Y578),这是 GIST 患者中最常见的突变之一,也是伊马替尼等酪氨酸激酶抑制剂的关键靶点。这使得 GIST-T1 成为研究 KIT 驱动的致癌作用和治疗反应的重要模型。 在长期培养中,GIST-T1细胞表现出稳定的增殖能力,并保持对伊马替尼的敏感性,除非经过专门筛选以获得耐药性。 为研究目的,已构建了GIST-T1的衍生耐药亚系,这些亚系表现出继发性KIT突变(如D820V或D820Y),从而能够研究耐药机制和适应性转录变化。这些耐药模型在与解毒、细胞周期调控及逃避免死相关的基因中均显示出改变。

GIST-T1还推动了GIST中新型致癌驱动因子的发现,包括在伊马替尼耐药亚系中鉴定的EXOC2-AK7等融合基因。功能研究表明,这些融合基因可增强GIST细胞的增殖和迁移能力,并使其对伊马替尼敏感,从而指明了新的治疗途径。 GIST相关超级增强子及转录因子网络(例如在转移进展中发挥作用的HAND1)的存在,进一步巩固了该模型在解析GIST表观遗传和转录架构方面的实用价值。总而言之,GIST-T1为研究胃肠道间质瘤的生物学特性、药物反应及耐药机制提供了一个经过遗传学和表型验证的可靠系统。

Tissue
转移性
Disease
胃肠道间质瘤
Metastatic site
胸腔积液
Synonyms
  • GIST-T-1
  • GISTT1
  • T1

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Cell type
卡哈尔间质细胞
Doubling time
48 小时

04Identifiers

Citation
GIST-T1(Cytion 产品编号 305777)
NCBI TaxID
9606
Cellosaurus
CVCL_4976

05Expression and susceptibility

Mutational profile
突变:KIT,简单型,p.Val560_Tyr578del (c.1679_1735del),杂合子

06Handling and culture

Culture medium
RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Seeding density
1 至 4 × 10⁴ 个细胞/cm²
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

Donor age
47 年
Ethnicity
日语