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

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

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

25 Downloads (Pure)

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
Article number103202
Number of pages18
JournalSTAR Protocols
Volume5
Issue number3
Early online date20 Jul 2024
DOIs
Publication statusPublished - 20 Sept 2024

Keywords

  • Cell Differentiation
  • Cell-based Assays
  • Stem Cells

Cite this