Experimentell Metastas och CTL adoptiv transfer Immunterapi musmodell
Summary
En experimentell lungmetastaser och CTL immunterapi musmodell för analys av tumörceller-T-cell interaktion<em> In vivo</em>.
Abstract
Experimentell metastaser musmodell är en enkel och ändå fysiologiskt relevanta metastaser modell. Tumörcellerna injiceras intravenöst (iv) i vener mus svans och kolonisera i lungorna, vilket, som liknar de sista stegen i tumörcellen spontana metastaser: överlevnad i cirkulationen, extravasering och kolonisation i distala organ. Från en terapeutisk synvinkel är den experimentella metastas modellen enklaste och ideala modellen eftersom målet av terapier är ofta slutpunkten för metastas: etablerade metastaserande tumör i distala orgel. I denna modell är tumörcellerna injiceras iv i mus svans vener och tillåtas att kolonisera och växa i lungorna. Tumör-specifik CTL sedan injiceras iv i metastaser bärande mus. Antalet och storleken på lungmetastaser kan styras av antalet tumörceller som ska injiceras och tidpunkten för tumörtillväxt. Därför är olika stadier av metastaser, från minimal metastaser till omfattande metastasering, kan modelleras. Lungmetastaser analyseras genom inflation med bläck, vilket gör det lättare att visuell observation och kvantifiering.
Protocol
Disclosures
The authors have nothing to disclose.
Acknowledgements
Finansierats med bidrag från National Institutes of Health (CA133085 till KL) och American Cancer Society (RSG-09-209-01-TBG till KL).
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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