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

doi:10.1016/j.xpro.2024.103202

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

School of Cellular and Molecular Medicine

Research output: Contribution to journal › Article (Academic Journal) › peer-review

Abstract

Leukemia 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 leukemia (PDX-ALL) cells with human mesenchymal stem cells (MSCs). 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.

For complete details on the use and execution of this protocol, please refer to Pal et al.1,2.

Original language	English
Article number	103202
Pages (from-to)	1-18
Number of pages	18
Journal	STAR Protocols
Volume	5
Issue number	3
Early online date	20 Jul 2024
DOIs	https://doi.org/10.1016/j.xpro.2024.103202
Publication status	Published - 20 Sept 2024

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© 2024 The Authors

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abstract = "Leukemia 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 leukemia (PDX-ALL) cells with human mesenchymal stem cells (MSCs). 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. For complete details on the use and execution of this protocol, please refer to Pal et al.1,2.",

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AU - Hockney, Sean

AU - Knill, Carly

AU - McDonald, David

AU - Filby, Andrew

AU - Pal, Deepali

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AB - Leukemia 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 leukemia (PDX-ALL) cells with human mesenchymal stem cells (MSCs). 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. For complete details on the use and execution of this protocol, please refer to Pal et al.1,2.

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