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HROG17 T1 M1
Tissue
大脑
Disease
胶质母细胞瘤

Documents

HROG17 T1 M1

Cell line · Product number 300875

HROG17 T1 M1 is a primary human glioblastoma multiforme (GBM) cell line established from a tumor specimen resected from an adult patient diagnosed with WH...

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01General information

Description

HROG17 T1 M1系由成人患者切除的肿瘤标本建立的人源原发性多形性胶质母细胞瘤(GBM)细胞系,该患者经诊断为WHO IV级胶质母细胞瘤。 标记"T1"表示样本取自首次手术时间点,"M1"则指由此肿瘤衍生的对应体外模型。该细胞系在HROG(汉萨城市罗斯托克胶质瘤)平台内建立,该平台专注于建立超低代次胶质瘤培养体系,以保留患者特异性分子及表型特征。

HROG17 T1 M1在标准培养条件下呈粘附生长,并展现原代胶质母细胞瘤培养物典型的成纤维细胞样形态。免疫表型分析表明,该细胞系表达胶质纤维酸性蛋白(GFAP)、巢蛋白和波形蛋白等胶质细胞及神经谱系相关标志物,符合高级别星形细胞瘤起源特征。 HROG细胞库的分子特征分析涵盖临床相关参数,包括MGMT启动子甲基化、EGFR扩增状态,以及TP53、IDH1/2、KRAS和BRAF等关键基因的突变分析,证实培养体系能保留肿瘤特异性基因组改变。

HROG17 T1 M1已被用于评估对胶质母细胞瘤标准治疗药物的敏感性,包括烷化剂类化疗药物及其他靶向化合物。 HROG模型间的比较分析表明,低代次培养物在早期代次中能保持稳定的形态、生长动力学及药物反应特征。作为患者来源的低代次胶质母细胞瘤模型,HROG17 T1 M1为研究高级别胶质瘤的肿瘤生物学特性、治疗反应及肿瘤间异质性提供了具有临床相关性的体外研究平台。

Tissue
大脑
Disease
胶质母细胞瘤

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
HROG17 T1 M1(Cytion 目录号 300875)
NCBI TaxID
9606
Cellosaurus
CVCL_B7FQ

04Handling and culture

Culture medium
DMEM:Ham's F12(1:1),w: 3.1 g/L 葡萄糖,w: 2.5 mM L-谷氨酰胺,w: 15 mM HEPES,w: 0.5 mM 丙酮酸钠,w: 1.2 g/L NaHCO3(Cytion 文章编号 820400a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
TrypLE Express,37°C,10 分钟、
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
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.

05Quality 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
作为冷冻保存培养基,我们使用 50% 基础培养基 + 40% FBS + 10% DMSO 或 CM-1(Cytion 目录号 800100),其中包括优化的渗透保护剂和代谢稳定剂,以促进恢复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。

06Donor

Donor age
70岁
Ethnicity
高加索人