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Isolering og karakterisering af exosomer fra skeletmuskelfibroblaster
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Isolering og karakterisering af exosomer fra skeletmuskelfibroblaster

Article DOI: 10.3791/61127-v • 06:27 min • May 16th, 2020
May 16th, 2020 •
1, 1, 1, 1
1Department of Genetics, St. Jude Children's Research Hospital

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Denne protokol illustrerer 1) isolering og dyrkning af primære fibroblaster fra den voksne muses gastrocnemiusmuskel samt 2) oprensning og karakterisering af exosomer ved hjælp af en differentiel ultracentrifugeringsmetode kombineret med saccharosedensitetsgradienter efterfulgt af western blot-analyser.

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Isolation karakterisering exosomer skeletmuskelfibroblaster ekstracellulære vesikler biologiske væsker ultracentrifugering ultrafiltrering størrelsesekskluderingskromatografi storskala eksosomoprensning primære fibroblaster voksne museskeletmuskler kulturmedier sekventielle centrifugeringstrin saccharosedensitetsgradienter exosomale præparater Western blot-analyser kanoniske markører (Alix CD9 CD81) elektronmikroskopi massespektrometri optagelseseksperimenter funktionelle Undersøgelser
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