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"In vitro" and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells.

Astori G, Vignati F, Bardelli S, Tubio M, Gola M, Albertini V, Bambi F, Scali G, Castelli D, Rasini V, Soldati G, Moccetti T - J Transl Med (2007)

Bottom Line: The CFU-F frequency in the SVF was 1/4880, a value about seven times greater than the data reported for bone marrow.AT-MSC were able to differentiate along the osteogenic adipogenic and chondrogenic lineages.The data here reported, further contribute to the characterization of SVF, a tissue providing an alternative as a source of MSC for clinical applications.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cell Therapy Unit, Cardiocentro Ticino, Via Tesserete 48, CH-6900 Lugano, Switzerland. giuseppe.astori@cardiocentro.org

ABSTRACT

Background: The stromal vascular fraction (SVF) is a heterogeneous cell population derived from the adipose tissue. There is still a lack of information concerning the characterization of the cell subpopulations constituting the SVF as well as its mesenchymal and haematopoietic potential. Furthermore there are great variations in its phenotypical characterization.

Methods: Composition of SVF was investigated by FACS analysis, cytological and "in vitro" assays. We studied CD34+ population by combining FACS with human CFC (colony-forming-cell haematopoietic assay). The endothelial fraction was investigated by quantifying the co-expression of specific markers (CD146, CD105, CD31 and UEA-1). Mesenchymal potential was assessed by CFU-F assay and cultured AT-MSC were characterized by a 5-color FACS analysis. The multipotent differentiation potential (osteogenic, adipogenic and chondrogenic) was investigated both at cellular and molecular level.

Results: We identified in the SVF two CD34+ populations with a marked difference in the intensity of antigen expression, the majority of the cells expressing CD34 at low intensity. Moreover, two CD146+ cell populations were clearly distinguishable in the SVF:a CD146 dim accounting for 9.9% of the total SVF cells and a CD146+ bright cell population accounting for about 39.3%. The frequency of CFC clones was comparable with the one reported for peripheral blood. Endothelial cells account for about 7.7% of the SVF cells. AT-MSC differenced in the osteogenic adipogenic and chondrogenic lineage.

Conclusion: The SVF is not a homogeneous cell population, and its final composition could be influenced both by the flow cytometric technique analysis and the SVF extraction steps. The CFU-F frequency in the SVF was 1/4880, a value about seven times greater than the data reported for bone marrow. The antigenic profile of AT-MSC was comparable with bone-marrow derived MSC. AT-MSC were able to differentiate along the osteogenic adipogenic and chondrogenic lineages. The data here reported, further contribute to the characterization of SVF, a tissue providing an alternative as a source of MSC for clinical applications.

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Endothelial cell markers expression on primary SVF cell population. The cells represented in the CD146 vs UEA-1 plot are CD45-.
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Figure 2: Endothelial cell markers expression on primary SVF cell population. The cells represented in the CD146 vs UEA-1 plot are CD45-.

Mentions: Endothelial cell-associated markers, including CD31 [20], CD105 [21] and CD146 are expressed on SVF cells. The CD146 [22], a cell marker expressed on mature endothelia, pericytes, endothelial progenitors cells (EPC) and at low intensity on a subset of activated T lymphocytes [23] stains 47.1 ± 3.2 of the cells (range 44.8–49.3) As can be observed in Figure 2, two distinct CD146 dim and CD146 bright cell clusters can be clearly distinguished. To finally confirm that mature endothelial cells are represented in the SVF, we have stained the cells in triplicate with CD45, CD146 and UEA-1. The results obtained suggest that it is possible to identify in the SVF a cell population having an endothelial phenotype defined as CD45-, CD146+ and UEA-1+ accounting for about the 7.7% of the total cells. (Figure 2).


"In vitro" and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells.

Astori G, Vignati F, Bardelli S, Tubio M, Gola M, Albertini V, Bambi F, Scali G, Castelli D, Rasini V, Soldati G, Moccetti T - J Transl Med (2007)

Endothelial cell markers expression on primary SVF cell population. The cells represented in the CD146 vs UEA-1 plot are CD45-.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2198906&req=5

Figure 2: Endothelial cell markers expression on primary SVF cell population. The cells represented in the CD146 vs UEA-1 plot are CD45-.
Mentions: Endothelial cell-associated markers, including CD31 [20], CD105 [21] and CD146 are expressed on SVF cells. The CD146 [22], a cell marker expressed on mature endothelia, pericytes, endothelial progenitors cells (EPC) and at low intensity on a subset of activated T lymphocytes [23] stains 47.1 ± 3.2 of the cells (range 44.8–49.3) As can be observed in Figure 2, two distinct CD146 dim and CD146 bright cell clusters can be clearly distinguished. To finally confirm that mature endothelial cells are represented in the SVF, we have stained the cells in triplicate with CD45, CD146 and UEA-1. The results obtained suggest that it is possible to identify in the SVF a cell population having an endothelial phenotype defined as CD45-, CD146+ and UEA-1+ accounting for about the 7.7% of the total cells. (Figure 2).

Bottom Line: The CFU-F frequency in the SVF was 1/4880, a value about seven times greater than the data reported for bone marrow.AT-MSC were able to differentiate along the osteogenic adipogenic and chondrogenic lineages.The data here reported, further contribute to the characterization of SVF, a tissue providing an alternative as a source of MSC for clinical applications.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cell Therapy Unit, Cardiocentro Ticino, Via Tesserete 48, CH-6900 Lugano, Switzerland. giuseppe.astori@cardiocentro.org

ABSTRACT

Background: The stromal vascular fraction (SVF) is a heterogeneous cell population derived from the adipose tissue. There is still a lack of information concerning the characterization of the cell subpopulations constituting the SVF as well as its mesenchymal and haematopoietic potential. Furthermore there are great variations in its phenotypical characterization.

Methods: Composition of SVF was investigated by FACS analysis, cytological and "in vitro" assays. We studied CD34+ population by combining FACS with human CFC (colony-forming-cell haematopoietic assay). The endothelial fraction was investigated by quantifying the co-expression of specific markers (CD146, CD105, CD31 and UEA-1). Mesenchymal potential was assessed by CFU-F assay and cultured AT-MSC were characterized by a 5-color FACS analysis. The multipotent differentiation potential (osteogenic, adipogenic and chondrogenic) was investigated both at cellular and molecular level.

Results: We identified in the SVF two CD34+ populations with a marked difference in the intensity of antigen expression, the majority of the cells expressing CD34 at low intensity. Moreover, two CD146+ cell populations were clearly distinguishable in the SVF:a CD146 dim accounting for 9.9% of the total SVF cells and a CD146+ bright cell population accounting for about 39.3%. The frequency of CFC clones was comparable with the one reported for peripheral blood. Endothelial cells account for about 7.7% of the SVF cells. AT-MSC differenced in the osteogenic adipogenic and chondrogenic lineage.

Conclusion: The SVF is not a homogeneous cell population, and its final composition could be influenced both by the flow cytometric technique analysis and the SVF extraction steps. The CFU-F frequency in the SVF was 1/4880, a value about seven times greater than the data reported for bone marrow. The antigenic profile of AT-MSC was comparable with bone-marrow derived MSC. AT-MSC were able to differentiate along the osteogenic adipogenic and chondrogenic lineages. The data here reported, further contribute to the characterization of SVF, a tissue providing an alternative as a source of MSC for clinical applications.

Show MeSH
Related in: MedlinePlus