| Autor: |
Hardy, W. Reef, Moldovan, Nicanor I., Moldovan, Leni, Livak, Kenneth J., Datta, Krishna, Goswami, Chirayu, Corselli, Mirko, Traktuev, Dmitry O., Murray, Iain R., Péault, Bruno, March, Keith |
| Zdroj: |
Stem Cells; May2017, Vol. 35 Issue 5, p1273-1289, 17p |
| Abstrakt: |
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. S tem C ells 2017;35:1273-1289 [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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