実験的転移とCTLの養子移入免疫療法のマウスモデル
Summary
腫瘍細胞- T細胞相互作用の分析のための実験的肺転移とCTL免疫療法のマウスモデル<em> in vivoで</em>。
Abstract
実験的転移のマウスモデルは、生理学的に関連はまだシンプルと転移モデルです。循環の生存率、遠位臓器の血管外漏出と植民地化：腫瘍細胞は腫瘍細胞自然転移の最後のステップに似て、それによって、マウスの尾静脈に（IV）静脈内に注射し、肺に植民地化されています。遠位臓器で確立された転移性腫瘍：治療の目標はしばしば転移のエンドポイントであるので、治療的観点から、実験的転移モデルは最も単純で理想的なモデルです。このモデルでは、腫瘍細胞は、マウスの尾静脈に静脈注射し、肺に植民地化と拡張されます。腫瘍特異的CTLは、その後、転移、担癌マウスに静脈注射されています。肺転移の数と大きさが注入される腫瘍細胞の数と腫瘍の成長の時間によって制御することができます。したがって、転移の様々な段階では、広範な転移への最小限の転移から、モデル化することができます。肺転移は、このように簡単に目視観察および定量を可能に、インクでインフレによって分析されています。
Protocol
1。実験的転移モデルマウス腫瘍細胞の注射の前日に、シードone T75フラスコ上で1 × 10 7〜急成長中の腫瘍細胞を得るために、10％血清を含むRPMI培地10mLに細胞をCMS4 – MET。 37℃で一晩インキュベート℃、 注射の日に、培地を除去し、PBSで細胞を1回洗い流した後、37℃で0.05％トリプシン- EDTAで腫瘍細胞を収穫℃で5分間。 10％血清を含むRPMI培地10mlで反応を停止します。コニ…
Disclosures
The authors have nothing to disclose.
Acknowledgements
国立衛生研究所（KLへCA133085）と米国癌学会からの補助金（KLにRSG – 09 – 209 – 01 – TBG）によってサポートされています。
Materials
Solutions:
India Ink Solution (17):
- Pour 150 ml of distilled water into a 250 ml flask.
- Add 4 drops ammonium hydroxide to the distilled water.
- Add 30 ml India Ink stock (i.e. Sanford Black Magic Waterproof Drawing Ink 4465 Item 44011) to the ammonia and water mixture.
- Top off with distilled water to a volume of 200 ml. Solution is ready for injection.
Fekete’s Solution (17):
Fekete’s solution is used to bleach India ink-inflated tumor-bearing lungs to distinguish white tumor nodules from the black background of normal tissues.
- Add 350ml 95% EtOH to 1L glass bottle.
- Add 150ml distilled water
- Add 50ml formaldehyde
- Add 25ml glacial acidic acid
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Tags
Experimental MetastasisMouse ModelCTL Adoptive Transfer ImmunotherapyTumor CellsIntravenous InjectionTail VeinsLungsMetastasis ModelTherapeutic Point Of ViewEstablished Metastatic TumorDistal OrganLung MetastasesTumor-specific CTLsInjectionColonization And GrowthNumber And Size ControlStages Of MetastasisMinimal MetastasisExtensive MetastasisInk Inflation Analysis
Cite This Article
Zimmerman, M., Hu, X., Liu, K. Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model. J. Vis. Exp. (45), e2077, doi:10.3791/2077 (2010).