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

被引：0
|
作者：
Parker, Jessica [1 ]
Hockney, Sean [1 ]
Knill, Carly [3 ,5 ]
McDonald, David [3 ,5 ]
Filby, Andrew [3 ,5 ]
Pal, Deepali [1 ,2 ,4 ]
机构：
[1] Northumbria Univ, Dept Appl Sci, Newcastle Upon Tyne NE1 8ST, Tyne & Wear, England
[2] Newcastle Univ, Fac Med Sci, Translat & Clin Res Inst, Wolfson Childhood Canc Res Ctr, Herschel Bldg Level 6,Brewery Lane, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England
[3] Newcastle Univ, Flow Cytometry Core Facil FCCF, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England
[4] Univ Bristol, Sch Cellular & Mol Med, Biomed Sci Bldg, Bristol BS8 1TD, Avon, England
[5] Newcastle Univ, Biosci Inst, Fac Med Sci, Innovat Methodol & Applicat IMA Res Theme, Newcastle Upon Tyne, Tyne & Wear, England
来源：
STAR PROTOCOLS | 2024年 / 5卷 / 03期
基金：
英国国家替代、减少和改良动物研究中心;
关键词：
D O I：
10.1016/j.xpro.2024.103202
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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 PDXALL 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 1,2
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页数：18
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