Experimentele metastase en CTL adoptieve overdracht Immunotherapie Muis Model
Summary
Een experimentele long metastase en CTL immunotherapie muis-model voor de analyse van tumorcellen-T-cel-interactie<em> In vivo</em>.
Abstract
Experimenteel metastase muismodel is een eenvoudige en toch fysiologisch relevante metastase model. De tumorcellen worden intraveneus geïnjecteerd (iv) in de muis staart aderen en koloniseren in de longen, waardoor, lijkt op de laatste stappen van tumorcellen spontane metastase: overleven in de bloedsomloop, extravasatie en kolonisatie in de distale organen. Vanuit therapeutisch oogpunt, de experimentele metastase model is de eenvoudigste en ideale model, omdat het doel van therapieën is vaak het eindpunt van metastase: opgericht gemetastaseerde tumor in de distale orgel. In dit model worden tumorcellen ingespoten iv in de muis staart aderen en toegestaan om te koloniseren en te groeien in de longen. Tumor-specifieke CTL's worden vervolgens geïnjecteerd in de uitzaaiingen iv-dragende muis. Het aantal en de grootte van de long metastasen kan worden gecontroleerd door het aantal tumorcellen te worden geïnjecteerd en de tijd van tumorgroei. Daarom verschillende stadia van metastase, van minimale metastase tot uitgebreide metastase, kan worden gemodelleerd. Longmetastasen worden geanalyseerd door de inflatie met inkt, waardoor makkelijker visuele waarneming en kwantificering.
Protocol
Disclosures
The authors have nothing to disclose.
Acknowledgements
Ondersteund door subsidies van de National Institutes of Health (CA133085 naar KL) en de American Cancer Society (RSG-09-209-01-TBG naar 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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