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During growth phase in vivo, tumor tissue xenografted in mouse model systems is vascularized and infiltrated by cells of murine origin. Such contamination makes molecular downstream analysis, like expression profiling or next-generation sequencing, challenging. In addition, culturing human tumor cells is frequently hampered by murine fibroblasts overgrowing target cells.
In this application protocol, we describe a time-saving workflow that overcomes these limitations and increases sensitivity of downstream analyses. Xenograft tumor tissue is dissociated into a viable single-cell suspension and untouched human tumor cells are isolated using MACS® technology.
Create a viable single-cell suspension from a solid xenograft tumor using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human. Follow the protocol from the kit data sheet.
Download data sheet
Human tumor cells are isolated by depleting the mouse cells using the Mouse Cell Depletion Kit and LS Columns. Follow the protocol from the kit data sheet.
Download the data sheet
We recommend filtering the magnetically labeled cell suspension to guarantee it is single-celled before separating it on the column.
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autoMACS, CliniMACS, the CliniMACS logo, CliniMACS Prodigy, CryoMACS, CytoMix, CytoStim, DendriMACS, ExiTron, ExpAct, FeraSpin, FeraTrack, GadoSpin, gentleMACS, LIFE 18, LIFE 21, MACS, the MACS logo, MACSductin, MACSelect, MACSfectin, MACSflex, MACSiBead, MACSiMAG, MACSmix, MACSprep, MACSQuant, MACSQuantify, MACSxpress, MidiMACS, MiniMACS, miRXplore, MultiMACS, NeuroBrew, NiraWave, OctoMACS, PepTivator, pMACS, PolySon, PrepProtect, QuadroMACS, REAfinity, REAlease, Rheo, StemMACS, StraightFrom, SuperAmp, SuperMACS, TexMACS, TheraSorb, thermoMACS, TransAct, Tyto, the Tyto logo, VarioMACS, Vio, Viobility, VioBlue, VioBright, VioGreen, Viscover, and µMACS are registered trademarks or trademarks of Miltenyi Biotec GmbH and/or its affiliates in various countries worldwide. Ficoll-Paque is a trademark of GE Healthcare companies. Vectofusin-1 is a registered trademark of Genethon. All other trademarks mentioned in this document are the property of their respective owners and are used for identification purposes only. Copyright © 2018 Miltenyi Biotec GmbH and/or its affiliates. All rights reserved.