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

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, i...

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Detalles Bibliográficos
Autores principales: Qing, Ying, Gao, Lei, Han, Li, Su, Rui, Chen, Jianjun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8209740/
https://www.ncbi.nlm.nih.gov/pubmed/34169295
http://dx.doi.org/10.1016/j.xpro.2021.100603
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author Qing, Ying
Gao, Lei
Han, Li
Su, Rui
Chen, Jianjun
author_facet Qing, Ying
Gao, Lei
Han, Li
Su, Rui
Chen, Jianjun
author_sort Qing, Ying
collection PubMed
description 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).
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spelling pubmed-82097402021-06-23 Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion Qing, Ying Gao, Lei Han, Li Su, Rui Chen, Jianjun STAR Protoc Protocol 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). Elsevier 2021-06-11 /pmc/articles/PMC8209740/ /pubmed/34169295 http://dx.doi.org/10.1016/j.xpro.2021.100603 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Qing, Ying
Gao, Lei
Han, Li
Su, Rui
Chen, Jianjun
Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion
title Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion
title_full Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion
title_fullStr Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion
title_full_unstemmed Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion
title_short Evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion
title_sort evaluation of glycolytic rates in human hematopoietic stem/progenitor cells after target gene depletion
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8209740/
https://www.ncbi.nlm.nih.gov/pubmed/34169295
http://dx.doi.org/10.1016/j.xpro.2021.100603
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