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

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).

SUBMITTER: Qing Y

PROVIDER: S-EPMC8209740 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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

Qing Ying Y Gao Lei L Han Li L Su Rui R Chen Jianjun J

STAR protocols 20210611 2

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. Th ...[more]

PMID: 34169295

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