Isolation and profiling of viable tumor cells from human ex vivo glioblastoma cultures through single-cell transcriptomics

Summary: Single-cell RNA-sequencing (scRNA-seq) is becoming a ubiquitous method in profiling the cellular transcriptomes of both malignant and non-malignant cells from the human brain. Here, we present a protocol to isolate viable tumor cells from human ex vivo glioblastoma cultures for single-cell...

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Bibliographic Details
Published in:STAR Protocols
Main Authors: Junyi Zhang, Jakob Straehle, Kevin Joseph, Nicolas Neidert, Simon Behringer, Jonathan Göldner, Andreas Vlachos, Marco Prinz, Christian Fung, Jürgen Beck, Oliver Schnell, Dieter Henrik Heiland, Vidhya M. Ravi
Format: Article
Language:English
Published: Elsevier 2023-09-01
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166723003507
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Summary:Summary: Single-cell RNA-sequencing (scRNA-seq) is becoming a ubiquitous method in profiling the cellular transcriptomes of both malignant and non-malignant cells from the human brain. Here, we present a protocol to isolate viable tumor cells from human ex vivo glioblastoma cultures for single-cell transcriptomic analysis. We describe steps including surgical tissue collection, sectioning, culturing, primary tumor cells inoculation, growth tracking, fluorescence-based cell sorting, and population-enriched scRNA-seq. This comprehensive methodology empowers in-depth understanding of brain tumor biology at the single-cell level.For complete details on the use and execution of this protocol, please refer to Ravi et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667