Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromes

Summary: Drug testing assays in hematopoietic stem and progenitor cells (HSPCs) are fundamental in biological studies of myelodysplastic syndromes (MDS) but have historically entailed a technical challenge. This protocol allows the efficient isolation of MDS HSPCs from bone marrow mononuclear cell f...

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Main Authors: Irene Ganan-Gomez, Karen Clise-Dwyer, Simona Colla
Format: Article
Language:English
Published: Elsevier 2022-12-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S266616672200644X
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author Irene Ganan-Gomez
Karen Clise-Dwyer
Simona Colla
author_facet Irene Ganan-Gomez
Karen Clise-Dwyer
Simona Colla
author_sort Irene Ganan-Gomez
collection DOAJ
description Summary: Drug testing assays in hematopoietic stem and progenitor cells (HSPCs) are fundamental in biological studies of myelodysplastic syndromes (MDS) but have historically entailed a technical challenge. This protocol allows the efficient isolation of MDS HSPCs from bone marrow mononuclear cell fractions and their culturing with the support of stromal cells for improved maintenance during drug testing. Lastly, specific steps are given to quantify surviving cells and assess changes in the HSPC hierarchies.For complete details on the use and execution of this protocol, please refer to Ganan-Gomez et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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spelling doaj.art-41d5d3c09aa140d1b729b74dfd7f47b32022-12-22T04:41:57ZengElsevierSTAR Protocols2666-16672022-12-0134101764Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromesIrene Ganan-Gomez0Karen Clise-Dwyer1Simona Colla2Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA; Corresponding authorDepartment of Hematopoietic Biology & Malignancy, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USASummary: Drug testing assays in hematopoietic stem and progenitor cells (HSPCs) are fundamental in biological studies of myelodysplastic syndromes (MDS) but have historically entailed a technical challenge. This protocol allows the efficient isolation of MDS HSPCs from bone marrow mononuclear cell fractions and their culturing with the support of stromal cells for improved maintenance during drug testing. Lastly, specific steps are given to quantify surviving cells and assess changes in the HSPC hierarchies.For complete details on the use and execution of this protocol, please refer to Ganan-Gomez et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672200644XCell BiologyCell cultureCell isolationFlow Cytometry/Mass CytometryCell separation/fractionationCancer
spellingShingle Irene Ganan-Gomez
Karen Clise-Dwyer
Simona Colla
Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromes
STAR Protocols
Cell Biology
Cell culture
Cell isolation
Flow Cytometry/Mass Cytometry
Cell separation/fractionation
Cancer
title Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromes
title_full Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromes
title_fullStr Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromes
title_full_unstemmed Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromes
title_short Isolation, culture, and immunophenotypic analysis of bone marrow HSPCs from patients with myelodysplastic syndromes
title_sort isolation culture and immunophenotypic analysis of bone marrow hspcs from patients with myelodysplastic syndromes
topic Cell Biology
Cell culture
Cell isolation
Flow Cytometry/Mass Cytometry
Cell separation/fractionation
Cancer
url http://www.sciencedirect.com/science/article/pii/S266616672200644X
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AT simonacolla isolationcultureandimmunophenotypicanalysisofbonemarrowhspcsfrompatientswithmyelodysplasticsyndromes