Isolation and characterization of circulating pro-vascular progenitor cell subsets from human whole blood samples

Daniella C Terenzi, Ehab Bakbak, Justin Z Trac, Mohammad Al-Omran, Adrian Quan, Hwee Teoh, Subodh Verma, David A Hess, Daniella C Terenzi, Ehab Bakbak, Justin Z Trac, Mohammad Al-Omran, Adrian Quan, Hwee Teoh, Subodh Verma, David A Hess

Abstract

The examination of circulating pro-vascular progenitor cell frequency and function is integral in understanding aberrant blood vessel homeostasis in individuals with cardiometabolic disease. Here, we outline the characterization of progenitor cell subsets from peripheral blood using high aldehyde dehydrogenase (ALDH) activity, an intracellular detoxification enzyme previously associated with pro-vascular progenitor cell status. Using this protocol, cells can be examined by flow cytometry for ALDH activity and lineage restricted cell surface markers simultaneously. For complete details on the use and execution of this protocol, please refer to Terenzi et al. (2019) and Hess et al. (2019, 2020).

Keywords: Cell biology; Cell isolation; Flow cytometry/mass cytometry; Stem cells.

Conflict of interest statement

H.T. reports receiving honorarium from Boehringer Ingelheim and writing fees for unrelated diabetes-related manuscripts from Merck and Servier. S.V. holds a Tier 1 Canada Research Chair in Cardiovascular Surgery and reports receiving research grants and/or speaking honoraria from Amarin, Amgen, AstraZeneca, Bayer, Boehringer Ingelheim, Bristol-Myers Squibb, Eli Lilly, EOCI Pharmacomm Ltd, HLS Therapeutics, Janssen, Merck, Novartis, Novo Nordisk, PhaseBio, Sanofi, Sun Pharmaceuticals, and the Toronto Knowledge Translation Working Group. He is the President of the Canadian Medical and Surgical Knowledge Translation Research Group, a federally incorporated not-for-profit physician organization.

© 2021 The Author(s).

Figures

Graphical abstract
Graphical abstract
Figure 1
Figure 1
High aldehyde dehydrogenase (ALDH) expression simulates progenitor cell differentiation
Figure 3
Figure 3
Visual representation of sample preparation
Figure 2
Figure 2
Mononuclear cell isolation and staining for ALDH activity and lineage restricted cell surface markers
Figure 4
Figure 4
Sample workflow of precursor cell analyses by flow cytometry

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