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

Cell Differentiation Cell-based Assays Stem Cells

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
20 Jul 2024
Historique:
received: 01 02 2024
revised: 30 04 2024
accepted: 25 06 2024
medline: 21 7 2024
pubmed: 21 7 2024
entrez: 21 7 2024
Statut: aheadofprint

Résumé

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.

Identifiants

DOI: 10.1016/j.xpro.2024.103202 PMID: 39033505
pubmed: 39033505
pii: S2666-1667(24)00367-8
doi: 10.1016/j.xpro.2024.103202
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

103202

Informations de copyright

Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Auteurs

Jessica Parker (J)

Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK.

Sean Hockney (S)

Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK.

Carly Knill (C)

Flow Cytometry Core Facility (FCCF), Newcastle University, Newcastle upon Tyne NE1 7RU, UK; Biosciences Institute, Innovation, Methodology and Application (IMA) Research Theme, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK.

David McDonald (D)

Flow Cytometry Core Facility (FCCF), Newcastle University, Newcastle upon Tyne NE1 7RU, UK; Biosciences Institute, Innovation, Methodology and Application (IMA) Research Theme, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK.

Andrew Filby (A)

Flow Cytometry Core Facility (FCCF), Newcastle University, Newcastle upon Tyne NE1 7RU, UK; Biosciences Institute, Innovation, Methodology and Application (IMA) Research Theme, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK.

Deepali Pal (D)

Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK; Wolfson Childhood Cancer Research Centre, Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Herschel Building Level 6, Brewery Lane, Newcastle upon Tyne NE1 7RU, UK; School of Cellular and Molecular Medicine, University of Bristol, Biomedical Sciences Building, University Walk, Bristol BS8 1TD, UK. Electronic address: deepali.pal@bristol.ac.uk.

Classifications MeSH