Abstract
Variation in numbers and functions of cells in fat tissues may affect therapeutic outcomes and adverse events after autologous fat tissue grafting in postmastectomy breast cancer patients; however, the relevant information regarding cellular components is still incomplete. Phenotypic characterization of heterogeneous cell subsets in stromal vascular fraction (SVF) isolated from fat tissues by flow cytometry was also limited to a combination of few molecules. This study, therefore, developed a polychromatic staining panel for an in-depth characterization of freshly isolated SVF and expanded adipose-derived stem cells (ADSC) from the patients. ADSC were found predominant in SVF (~65% of CD45− cells) with a homogenous phenotype of CD13+CD31−CD34+CD45−CD73+CD90+CD105−CD146− (~94% of total ADSC). Endothelial progenitor cells (EPC) and pericytes were minor (~18% and ~11% of CD45− cells, respectively) with large heterogeneity. Downregulation of CD34 and upregulation of CD105 in ADSC were profound at passage 3, showing a phenotype similar to the classical mesenchymal stem cells from the bone marrow. Results from this study demonstrated that fat tissue collected from patients contains ADSC with a highly homogenous phenotype. The in vitro culture of these cells maintained their homogeneity with modified CD34 and CD105 expression, suggesting the expansion from a single population of ADSC.
Highlights
White adipose tissue has been recognized as the alternative source for stromal precursors and stem cells
Our results showed that adipose-derived stem cells (ADSC), Endothelial progenitor cells (EPC), and pericytes were able to be distinguished by a minimum of 4 markers including CD31, CD34, CD45, and CD146
In our freshly isolated stromal vascular fraction (SVF), ADSC were the most abundant followed by EPC and pericytes (64:6 ± 14:2%, 17:8 ± 10:4%, and 11:2 ± 7:8% of CD45- cells, respectively) which differed from a finding by Agostini et al showing ADSC with 58:8 ± 16:6% and EPC with 43:2 ± 16:6% of CD34+CD45cells [27]
Summary
White adipose tissue has been recognized as the alternative source for stromal precursors and stem cells. White adipose tissue contains a single lipid droplet creating white to yellow appearance and functions by storing lipids for excessive energy, whereas brown adipose tissue comprises multiple small vacuoles with abundance of iron-containing mitochondria generating brown color and works through lipid burning for heat production [1,2,3]. White adipose tissue is found predominantly in subcutaneous and several visceral depots (e.g., abdomen, hip, and thigh); it becomes a sensible source for progenitor stem cells. Compared to the bone marrow—another recommended source of stem cells, the yield of mesenchymal stem cells (MSC) from white adipose tissue was able to reach 0.5–1:25 × 106 cells/gram adipose tissue [5, 6] while only
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