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

Research output: Contribution to journal › Article › peer-review

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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 language	English
Article number	103202
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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Cell Differentiation
Cell-based Assays
Stem Cells

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10.1016/j.xpro.2024.103202Licence: CC BY

Final manuscript 2406Accepted author manuscript, 1.45 MBLicence: CC BY
1-s2.0-S2666166724003678-mainFinal published version, 6.09 MBLicence: CC BY

Cite this

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T1 - Protocol for in vitro co-culture, proliferation, and cell cycle analyses of patient-derived leukemia cells

AU - Parker, Jessica

AU - Hockney, Sean

AU - Knill, Carly

AU - McDonald, David

AU - Filby, Andrew

AU - Pal, Deepali

PY - 2024/9/20

Y1 - 2024/9/20

N2 - 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.

AB - 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.

KW - Cell Differentiation

KW - Cell-based Assays

KW - Stem Cells

UR - https://www.scopus.com/pages/publications/85199128384

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DO - 10.1016/j.xpro.2024.103202

M3 - Article

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JF - STAR Protocols

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M1 - 103202

ER -

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

Abstract

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Keywords

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