Work

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

Year: 2021

Type: article

Abstract: 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).

Source: STAR Protocols

DOI: 10.1016/j.xpro.2021.100603

Authors: Ying Qing, Lei Gao, Li Han, Rui Su, Jianjun Chen

Institutions: City Of Hope National Medical Center, City of Hope, Beckman Research Institute

Language: English

Topic: Cancer, Hypoxia, and Metabolism

Subfield: Cancer Research

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,500