胃腸道間質(zhì)瘤-T1培養(yǎng)套裝

◆背景

胃腸道間質(zhì)瘤（GISTs）不像大多數(shù)的腫瘤生長在小腸和食道胃腸道，而是生長在胃粘膜下。間質(zhì)瘤產(chǎn)生于Cajal間質(zhì)細胞和腸道起搏細胞（pacemakercells）。

GIST-T1細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. ），從日本婦女胃部GISTs（胃腸道間質(zhì)瘤）中分離得到的細胞系。 

 

◆培養(yǎng)套裝的組成

●　GIST-T1（凍存細胞）1.0×10⁶ cells/vial

●　培養(yǎng)基　　250 mL

* 培養(yǎng)基的組成: DMEM，F(xiàn)BS，抗生素等。

 

◆基本信息

生物體: 人類
組織: 胃
培養(yǎng): 粘附性能

生物安全性: 等級1

性別: 女性
種族:亞洲
病毒檢測: HIV-1(-), HTLV-1(-), HBV(-), HCV(-), T.pallidum(-)

質(zhì)量檢查: 支原體（ – ）   

◆注意事項

●　因為細胞是來自人類組織，在研究實驗中，一定要穿戴手套和防護眼鏡。

●　從干冰包裝中取出凍存管，并立即放入液氮中儲存?zhèn)溆谩?br />

●　基于四國和高知大學的技術網(wǎng)絡許可協(xié)議（the license agreement ofTechno network Shikoku and 

　　Kochi University），GIST-T1細胞禁止提供（分發(fā)，出借，轉(zhuǎn)讓，許可等）給第三方。

  

購買 “GIST‐T1 細胞”協(xié)議

請遵守以下指示:

(1)　 胃腸道間質(zhì)瘤-T1培養(yǎng)套裝僅供研究使用。

(2)　 該產(chǎn)品僅支持項目的第三方使用，產(chǎn)品使用完后必須進行回收或銷毀。

(3)　我們禁止將GIST-T1培養(yǎng)套裝分發(fā)、出借、轉(zhuǎn)讓、許可、給超出了我們的控制或職責范圍的第三方。 

　　　 　 請下載并打印協(xié)議格式（PDF文件），并且在下訂單時傳真或電子郵件發(fā)送該協(xié)議給我們。

 

◆實驗示例

圖一  免疫組化和相差顯微鏡觀察

注意: 請使用推薦培養(yǎng)基（Cat.no＃PMC-GISTM-COS）對GIST-T1進行培養(yǎng)。

        使用其他培養(yǎng)基我們將不能保證實驗結(jié)果。

        Cosmo Bio無法保證客戶的實驗室凍存細胞。

 

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.