Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion

STAR Protocols

Rui Su¹  Lei Gao²  Li Han³  Jianjun Chen⁴  Ying Qing⁴ 

[1] Corresponding author;;Irell &Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA;Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA;

关键词: Cell isolation;    Cell-based Assays;    Cancer;    Metabolism;    Molecular Biology;    Gene Expression;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: This protocol describes how to evaluate cellular metabolic state, including the glycolytic activity, in live CD34+ hematopoietic stem/progenitor cells (HSPCs) upon depletion of a specific target gene. We detail the procedure of enriching mononuclear cells from human umbilical cord blood samples, isolation of CD34+ HSPCs using positive immunomagnetic separation, and the analysis of glycolytic activity in lentivirally transduced CD34+ HSPCs using Seahorse Assay. This protocol can be applied to evaluate potential aerobic glycolysis gene targets for cancer therapy.For complete details on the use and execution of this protocol, please refer to Qing et al. (2021).

【 授权许可】

Unknown