胃肠道间质瘤-T1培养套装

◆背景

胃肠道间质瘤（GISTs）不像大多数的肿瘤生长在小肠和食道胃肠道，而是生长在胃粘膜下。间质瘤产生于 Cajal 间质细胞和肠道起搏细胞（pacemakercells）。

GIST-T1 细胞系是田口尚弘副教授（Takahiro Taguchi; associate professor, Graduate School of Integrated Arts and Sciences, Kochi-University, Kochi, Japan. ），从日本妇女胃部 GISTs（胃肠道间质瘤）中分离得到的细胞系。 

◆培养套装的组成

●　GIST-T1（冻存细胞）1.0×10⁶ cells/vial

●　培养基　　250 mL

* 培养基的组成：DMEM，FBS，抗生素等。

◆基本信息

生物体：人类
组织：胃
培养：粘附性能

生物安全性：等级１

性别：女性
种族：亚洲
病毒检测：HIV-1(-), HTLV-1(-), HBV(-), HCV(-), T.pallidum(-)

质量检查：支原体（ – ）   

◆注意事项

●　因为细胞是来自人类组织，在研究实验中，一定要穿戴手套和防护眼镜。

●　从干冰包装中取出冻存管，并立即放入液氮中储存备用。

●　基于四国和高知大学的技术网络许可协议（the license agreement ofTechno network Shikoku and 

　　Kochi University），GIST-T1 细胞禁止提供（分发，出借，转让，许可等）给第三方。

购买 “GIST‐T1 细胞”协议

请遵守以下指示：

(1)　 胃肠道间质瘤-T1 培养套装仅供研究使用。

(2)　 该产品仅支持项目的第三方使用，产品使用完后必须进行回收或销毁。

(3)　我们禁止将 GIST-T1 培养套装分发、出借、转让、许可、给超出了我们的控制或职责范围的第三方。 

　　　 　 请下载并打印协议格式（PDF文件），并且在下订单时传真或电子邮件发送该协议给我们。

◆实验示例

图一  免疫组化和相差显微镜观察

注意: 请使用推荐培养基（Cat.no＃PMC-GISTM-COS）对 GIST-T1 进行培养。

        使用其他培养基我们将不能保证实验结果。

        Cosmo Bio 无法保证客户的实验室冻存细胞。

Protocol

A) Thawing of Cells 

1) 　Prepare a 100mm dish (Note: 100mm dish is recommended).

2)　 Warm culture medium to 37°C. 

3) 　Prepare a conical tube (for 15mL) added 10mL of culture medium. 

4) 　Carefully remove the cryovial from liquid nitrogen and thaw cells in a water bath at 37°C for 90 seconds. 

5)　 Transfer the cryovial into a laminar flow hood. Before opening, wipe the outside of the vial with 70% ethanol. 

6)　 Gently transfer the thawed cell suspension (1mL) into 10 mL of culture medium. 

7) 　Transfer 1mL of culture medium in the same conical tube back to the cryovial and pour the contents back to 15mL conical 

        tube. 

8) 　Centrifuge the cell suspension at approximately 200 ×g for 5 minutes at 4°C. 

9) 　Aspirate the supernatant without disrupting the pellet and resuspend the cells in 10mL of culture medium. 

10)  Transfer the cell suspension to 100mm dish and incubate the cells in 37°C, 5% CO2 incubator. 

11)  Replace the medium with fresh pre-warmed culture medium every 2 to 3 days.

B) Subculturing Note: Allow culture medium, HBSS(or PBS(-)), and 0.25% Trypsin to come to room temperature before use.

1) 　When the cells reach 70 -90% of confluent, they should be subcultured. 

2)　 Aspirate the medium. Rinse the dish with 10mL of HBSS or PBS (-). 

3)　 Add 1mL of 0.25% Trypsin, then incubate at 37°C for 4-6 minutes. 

4) 　Add 10mL of culture medium and disperse the cells with gentle pipetting. 

5)　 Transfer the cell suspension to conical tube and centrifuge at 200 ×g for 5 minutes at 4°C. 

6)　 Aspirate the supernatant without disrupting the pellet and resuspend the cells in 10mL of culture medium. 

7)　 Dilute the cell suspension by adding culture medium. A subcultivation ratio of 1:6 to 1:8 is recommended. 

8) 　Transfer the cell suspension to new 100mm dish and Incubate the cells in 37°C, 5% CO₂ incubator. 

9) 　Replace the medium with fresh pre-warmed culture medium every 2 to 3 days. 

10)   Culture the cells until the required density (70 -90% of confluent; Fig 1, C) is reached.

CSR_GIST-T1CultureKit.pdf

[1] Takahiro Taguchi, Hiroshi Sonobe, and Kazunari Yuri. et al. Conventional and Molecular Cytogenetic Characterization of a New Human Cell Line, GIST-T1, Established from Gastrointestinal Stromal Tumor. Lab Invest. 2002 May;82(5):663-5.