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

Irene Ganan-Gomez; Karen Clise-Dwyer; Simona Colla

doi:10.1016/j.xpro.2022.101764

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

STAR Protoc. 2022 Dec 16;3(4):101764. doi: 10.1016/j.xpro.2022.101764. Epub 2022 Oct 13.

Authors

Irene Ganan-Gomez¹, Karen Clise-Dwyer², Simona Colla³

Affiliations

¹ Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. Electronic address: iganan@mdanderson.org.
² Department of Hematopoietic Biology & Malignancy, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
³ Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.

Abstract

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).

Keywords: Cancer; Cell Biology; Cell culture; Cell isolation; Cell separation/fractionation; Flow Cytometry/Mass Cytometry; Stem Cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Bone Marrow Cells
Bone Marrow*
Hematopoietic Stem Cells
Humans
Myelodysplastic Syndromes*
Stromal Cells

Abstract

Publication types

MeSH terms

Grants and funding