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

Lookup NU author(s): Sean Hockney, Carly Knill, Dr David McDonald, Professor Andrew FilbyORCiD, Dr Deepali Pal

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

© 2024 The Authors. 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.

Publication metadata

Author(s): Parker J, Hockney S, Knill C, McDonald D, Filby A, Pal D

Publication type: Article

Publication status: Published

Journal: STAR Protocols

Year: 2024

Volume: 5

Issue: 3

Print publication date: 20/09/2024

Online publication date: 20/07/2024

Acceptance date: 02/04/2018

Date deposited: 29/07/2024

ISSN (electronic): 2666-1667

Publisher: Cell Press

URL: https://doi.org/10.1016/j.xpro.2024.103202

DOI: 10.1016/j.xpro.2024.103202

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