Protocol for in vitro co-culture, proliferation and cell cycle analyses of patient-derivedleukaemia cells

Jessica Parker, Sean Hockney, Carly Knill, David McDonald, Andrew Filby, Deepali Pal*

*Corresponding author for this work

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Abstract

Leukaemia niche impacts quiescence, however culturing patient derived samples ex vivo is technically challenging. Here, we present a protocol for in vitro co-culture of patient-derived-xenograft acute lymphoblastic leukaemia (PDX-ALL) cells with human mesenchymal stem cells (MSC). We describe steps for labeling PDX-ALL cells with CellTrace™ Violet dye to demonstrate MSC-primed PDX-ALL cycling. We then detail procedures to identify MSC-primed G0/quiescent PDX-ALL cells via Hoechst-33342 / Pyronin Y live cell cycle analysis.
Original languageEnglish
JournalSTAR Protocols
Publication statusAccepted/In press - 25 Jun 2024

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