Transcriptional Networks in Single Perivascular Cells Sorted from Human Adipose Tissue Reveal a Hierarchy of Mesenchymal Stem Cells

W Reef Hardy; Nicanor I Moldovan; Leni Moldovan; Kenneth J Livak; Krishna Datta; Chirayu Goswami; Mirko Corselli; Dmitry O Traktuev; Iain R Murray; Bruno Péault; Keith March

doi:10.1002/stem.2599

Transcriptional Networks in Single Perivascular Cells Sorted from Human Adipose Tissue Reveal a Hierarchy of Mesenchymal Stem Cells

Stem Cells. 2017 May;35(5):1273-1289. doi: 10.1002/stem.2599. Epub 2017 Mar 19.

Authors

W Reef Hardy^{1

2}, Nicanor I Moldovan³, Leni Moldovan³, Kenneth J Livak⁴, Krishna Datta⁴, Chirayu Goswami⁵, Mirko Corselli^{1

6}, Dmitry O Traktuev², Iain R Murray^{1

7}, Bruno Péault^{1

7}, Keith March²

Affiliations

¹ Department of Orthopaedic Surgery and Broad Stem Cell Center, University of California at Los Angeles, California, USA.
² Department of Medicine, University of Indiana, Indianapolis, Indiana, USA.
³ Department of Ophthalmology, IUPUI, Indianapolis, Indiana, USA.
⁴ Fluidigm Corporation, South San Francisco, California, USA.
⁵ Thomas Jefferson University Hospitals, Philadelphia, Pennsylvania, USA.
⁶ BD Biosciences, San Diego, California.
⁷ MRC Centre for Regenerative Medicine, University of Edinburgh, Scotland, United Kingdom.

PMID: 28233376
DOI: 10.1002/stem.2599

Abstract

Adipose tissue is a rich source of multipotent mesenchymal stem-like cells, located in the perivascular niche. Based on their surface markers, these have been assigned to two main categories: CD31^- /CD45^- /CD34⁺ /CD146^- cells (adventitial stromal/stem cells [ASCs]) and CD31^- /CD45^- /CD34^- /CD146⁺ cells (pericytes [PCs]). These populations display heterogeneity of unknown significance. We hypothesized that aldehyde dehydrogenase (ALDH) activity, a functional marker of primitivity, could help to better define ASC and PC subclasses. To this end, the stromal vascular fraction from a human lipoaspirate was simultaneously stained with fluorescent antibodies to CD31, CD45, CD34, and CD146 antigens and the ALDH substrate Aldefluor, then sorted by fluorescence-activated cell sorting. Individual ASCs (n = 67) and PCs (n = 73) selected from the extremities of the ALDH-staining spectrum were transcriptionally profiled by Fluidigm single-cell quantitative polymerase chain reaction for a predefined set (n = 429) of marker genes. To these single-cell data, we applied differential expression and principal component and clustering analysis, as well as an original gene coexpression network reconstruction algorithm. Despite the stochasticity at the single-cell level, covariation of gene expression analysis yielded multiple network connectivity parameters suggesting that these perivascular progenitor cell subclasses possess the following order of maturity: (a) ALDH^br ASC (most primitive); (b) ALDH^dim ASC; (c) ALDH^br PC; (d) ALDH^dim PC (least primitive). This order was independently supported by specific combinations of class-specific expressed genes and further confirmed by the analysis of associated signaling pathways. In conclusion, single-cell transcriptional analysis of four populations isolated from fat by surface markers and enzyme activity suggests a developmental hierarchy among perivascular mesenchymal stem cells supported by markers and coexpression networks. Stem Cells 2017;35:1273-1289.

Keywords: Adipose tissue; Mesenchymal stem cells; Pericyte; Single cell transcriptome; Tunica adventitia.

MeSH terms

Adipose Tissue / cytology*
Aldehyde Dehydrogenase / metabolism
Cell Differentiation / genetics
Cell Lineage*
Female
Flow Cytometry
Gene Expression Regulation
Gene Regulatory Networks*
Humans
Mesenchymal Stem Cells / cytology*
Mesenchymal Stem Cells / metabolism*
Middle Aged
Pericytes / cytology
Single-Cell Analysis

Substances

Aldehyde Dehydrogenase

Abstract

MeSH terms

Substances

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