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

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29 Scopus citations

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) ALDHbrASC (most primitive); (b) ALDHdimASC; (c) ALDHbrPC; (d) ALDHdimPC (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.

Original languageEnglish (US)
Pages (from-to)1273-1289
Number of pages17
JournalSTEM CELLS
Volume35
Issue number5
DOIs
StatePublished - May 2017

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Keywords

  • Adipose tissue
  • Mesenchymal stem cells
  • Pericyte
  • Single cell transcriptome
  • Tunica adventitia

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

Cite this

Hardy, W. R., Moldovan, N. I., Moldovan, L., Livak, K. J., Datta, K., Goswami, C., Corselli, M., Traktuev, D. O., Murray, I. R., Péault, B., & March, K. (2017). Transcriptional Networks in Single Perivascular Cells Sorted from Human Adipose Tissue Reveal a Hierarchy of Mesenchymal Stem Cells. STEM CELLS, 35(5), 1273-1289. https://doi.org/10.1002/stem.2599