Journal
CYTOMETRY PART A
Volume 83A, Issue 1, Pages 134-140Publisher
WILEY
DOI: 10.1002/cyto.a.22227
Keywords
multiparameter flow cytometry; adipose stromal vascular fraction; mesenchymal stem cells; endothelial progenitor cells; perivascular cells; pericytes; immunofluorescent microscopy; supra-adventitial adipose stromal cells
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Funding
- Diana Napper from The Glimmer of Hope Foundation
- NATIONAL CANCER INSTITUTE [R01CA114246, P30CA047904] Funding Source: NIH RePORTER
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The stromal-vascular fraction (SVF) of adipose tissue is a rich source of multipotent stem cells. We and others have described three major populations of stem/progenitor cells in this fraction, all closely associated with small blood vessels: endothelial progenitor cells (EPC, CD45-/CD31+/CD34+), pericytes (CD45-/CD31-/CD146+), and supra-adventitial adipose stromal cells (SA-ASC, CD45-/CD31-/CD146-/CD34+). EPC are luminal, pericytes are adventitial, and SA-ASC surround the vessel like a sheath. The multipotency of the pericytes and SA-ASC compartments is strikingly similar to that of CD45-/CD34-/CD73+/CD105+/CD90+ bone marrow-derived mesenchymal stem cells (BM-MSC). Here, we determine the extent to which this mesenchymal pattern is expressed on the three adipose stem/progenitor populations. Eight independent adipose tissue samples were analyzed in a single tube (CD105-FITC/CD73-PE/CD146-PETXR/ CD14-PECY5/CD33-PECY5/CD235A-PECY5/CD31-PECY7/CD90-APC/CD34-A700/ CD45-APCCY7/DAPI). Adipose EPC were highly proliferative with (14.3 +/- 2.8)% (mean +/- SEM) having > 2N DNA. About half (53.1 +/- 7.6)% coexpressed CD73 and CD105, and (71.9 +/- 7.4)% expressed CD90. Pericytes were less proliferative [(8.2 +/- 3.4)% > 2N DNA)] with a smaller proportion [(29.6 +/- 6.9)% CD73+/CD105+, (60.5 perpendicular to 10.2)% CD90+] expressing mesenchymal associated markers. However, the CD34+ subset of CD146+ pericytes were both highly proliferative [(15.1 +/- 3.6)% with > 2N DNA] and of uniform mesenchymal phenotype [(93.3 +/- 3.7)% CD73+/CD105+, (97.8 perpendicular to 0.7)% CD90+], suggesting transit amplifying progenitor cells. SA-ASC were the least proliferative [(3.7 +/- 0.8)%> 2N DNA] but were also highly mesenchymal in phenotype [(94.4 +/- 3.2)% CD73+/CD105+, (95.5 +/- 1.2)% CD901]. These data imply a progenitor/ progeny relationship between pericytes and SA-ASC, the most mesenchymal of SVF cells. Despite phenotypic and functional similarities to BM-MSC, SA-ASC are distinguished by CD34 expression. (C) 2012 International Society for Advancement of Cytometry
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