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Adipose Stem Cells Display Higher Regenerative Capacities and More Adaptable Electro-Kinetic Properties Compared to Bone Marrow-Derived Mesenchymal Stromal Cells

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ABSTRACT

Adipose stem cells (ASCs) have recently emerged as a more viable source for clinical applications, compared to bone-marrow mesenchymal stromal cells (BM-MSCs) because of their abundance and easy access. In this study we evaluated the regenerative potency of ASCs compared to BM-MSCs. Furthermore, we compared the dielectric and electro-kinetic properties of both types of cells using a novel Dielectrophoresis (DEP) microfluidic platform based on a printed circuit board (PCB) technology. Our data show that ASCs were more effective than BM-MSCs in promoting neovascularization in an animal model of hind-limb ischemia. When compared to BM-MSCs, ASCs displayed higher resistance to hypoxia-induced apoptosis, and to oxidative stress-induced senescence, and showed more potent proangiogenic activity. mRNA expression analysis showed that ASCs had a higher expression of Oct4 and VEGF than BM-MSCs. Furthermore, ASCs showed a remarkably higher telomerase activity. Analysis of the electro-kinetic properties showed that ASCs displayed different traveling wave velocity and rotational speed compared to BM-MSCs. Interestingly, ASCs seem to develop an adaptive response when exposed to repeated electric field stimulation. These data provide new insights into the physiology of ASCs, and evidence to their potential superior potency compared to marrow MSCs as a source of stem cells.

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Representative histological analysis of hind limb muscles: Gastrocnemius muscles were collected after 4 weeks of cell therapy.Tissue samples were stained with: (a) H & E showing muscle degeneration in the ischemic control group and infiltration of lymphocytes (*) compared to normal looking muscles in the BM-MSCs and ASCs treated groups (b) Positive staining for-CD31, in transplanted mice, especially In the ASCs-transplanted group (c) CD34 expression is pronounced in the BM-MSC-transplanted group (d) Increased expression of VEGF especially in the ASC-treated group (e) Staining with anti-αSMA is more pronounced in the ASCs group (f) staining of both tissues with anti-MMP9. Quantitative evaluation of the expression levels of CD31 (g), CD34 (h) and αSMA (i) was evaluated by counting the number of positive cells in each group. Data are shown as mean ± S.D. (error bars). Scale bars, 200 μm.
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f1: Representative histological analysis of hind limb muscles: Gastrocnemius muscles were collected after 4 weeks of cell therapy.Tissue samples were stained with: (a) H & E showing muscle degeneration in the ischemic control group and infiltration of lymphocytes (*) compared to normal looking muscles in the BM-MSCs and ASCs treated groups (b) Positive staining for-CD31, in transplanted mice, especially In the ASCs-transplanted group (c) CD34 expression is pronounced in the BM-MSC-transplanted group (d) Increased expression of VEGF especially in the ASC-treated group (e) Staining with anti-αSMA is more pronounced in the ASCs group (f) staining of both tissues with anti-MMP9. Quantitative evaluation of the expression levels of CD31 (g), CD34 (h) and αSMA (i) was evaluated by counting the number of positive cells in each group. Data are shown as mean ± S.D. (error bars). Scale bars, 200 μm.

Mentions: BM-MSCs and ASCs were characterized by their cell surface marker expression using flow cytometry and by their adipogenic and osteogenic differentiation potential (Supplemental Fig. 1B & C). Both BM-MSCs and ASCs were shown to be positive for CD29, CD90 and were negative to CD45 surface antigens (Supplemental Fig. 1D). This expression profile is in accordance with the International Society for Cellular Therapy Statement of minimal criteria for defining MSC31. To compare the in vivo differences between BM-MSCs and ASCs in promoting angiogenesis in an animal model of hind limb ischemia, the gastrocnemius muscles were collected 3 weeks after administration of either ASCs, or BM-MSCs. H & E staining showed muscle degeneration and lymphocyte infiltration in the ischemic control group while muscles in limbs treated with both BM-MSCs as well as ASCs were protected after cell transplantation (Fig. 1a). Immunohistological staining for CD31 and CD34 antigens showed increase of the number cells expressing these antigens (endothelial cells and endothelial progenitor cells respectively) in the ASC-treated group and the BM-MSC-treated group, respectively. (Fig. 1b and c). On the other hand, VEGF expression was especially prominent in the ASC-treated group (Fig. 1d). Immunostaining for αSMA, a marker of vascular smooth muscle cells, and MMP9, which is essential for neovascularization and initiating angiogenesis was higher in the ASC-transplanted group (Fig. 1e and f). The expression of CD31, CD34 and αSMA was quantified by counting the number of positive cells (Fig. 1g, h and i). Representative histological analysis of original and magnified images of hind limb muscles stained for CD31, CD34, VEGF, αSMA and MMP9 are shown in Supplemental Figures 2–6.


Adipose Stem Cells Display Higher Regenerative Capacities and More Adaptable Electro-Kinetic Properties Compared to Bone Marrow-Derived Mesenchymal Stromal Cells
Representative histological analysis of hind limb muscles: Gastrocnemius muscles were collected after 4 weeks of cell therapy.Tissue samples were stained with: (a) H & E showing muscle degeneration in the ischemic control group and infiltration of lymphocytes (*) compared to normal looking muscles in the BM-MSCs and ASCs treated groups (b) Positive staining for-CD31, in transplanted mice, especially In the ASCs-transplanted group (c) CD34 expression is pronounced in the BM-MSC-transplanted group (d) Increased expression of VEGF especially in the ASC-treated group (e) Staining with anti-αSMA is more pronounced in the ASCs group (f) staining of both tissues with anti-MMP9. Quantitative evaluation of the expression levels of CD31 (g), CD34 (h) and αSMA (i) was evaluated by counting the number of positive cells in each group. Data are shown as mean ± S.D. (error bars). Scale bars, 200 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5121630&req=5

f1: Representative histological analysis of hind limb muscles: Gastrocnemius muscles were collected after 4 weeks of cell therapy.Tissue samples were stained with: (a) H & E showing muscle degeneration in the ischemic control group and infiltration of lymphocytes (*) compared to normal looking muscles in the BM-MSCs and ASCs treated groups (b) Positive staining for-CD31, in transplanted mice, especially In the ASCs-transplanted group (c) CD34 expression is pronounced in the BM-MSC-transplanted group (d) Increased expression of VEGF especially in the ASC-treated group (e) Staining with anti-αSMA is more pronounced in the ASCs group (f) staining of both tissues with anti-MMP9. Quantitative evaluation of the expression levels of CD31 (g), CD34 (h) and αSMA (i) was evaluated by counting the number of positive cells in each group. Data are shown as mean ± S.D. (error bars). Scale bars, 200 μm.
Mentions: BM-MSCs and ASCs were characterized by their cell surface marker expression using flow cytometry and by their adipogenic and osteogenic differentiation potential (Supplemental Fig. 1B & C). Both BM-MSCs and ASCs were shown to be positive for CD29, CD90 and were negative to CD45 surface antigens (Supplemental Fig. 1D). This expression profile is in accordance with the International Society for Cellular Therapy Statement of minimal criteria for defining MSC31. To compare the in vivo differences between BM-MSCs and ASCs in promoting angiogenesis in an animal model of hind limb ischemia, the gastrocnemius muscles were collected 3 weeks after administration of either ASCs, or BM-MSCs. H & E staining showed muscle degeneration and lymphocyte infiltration in the ischemic control group while muscles in limbs treated with both BM-MSCs as well as ASCs were protected after cell transplantation (Fig. 1a). Immunohistological staining for CD31 and CD34 antigens showed increase of the number cells expressing these antigens (endothelial cells and endothelial progenitor cells respectively) in the ASC-treated group and the BM-MSC-treated group, respectively. (Fig. 1b and c). On the other hand, VEGF expression was especially prominent in the ASC-treated group (Fig. 1d). Immunostaining for αSMA, a marker of vascular smooth muscle cells, and MMP9, which is essential for neovascularization and initiating angiogenesis was higher in the ASC-transplanted group (Fig. 1e and f). The expression of CD31, CD34 and αSMA was quantified by counting the number of positive cells (Fig. 1g, h and i). Representative histological analysis of original and magnified images of hind limb muscles stained for CD31, CD34, VEGF, αSMA and MMP9 are shown in Supplemental Figures 2–6.

View Article: PubMed Central - PubMed

ABSTRACT

Adipose stem cells (ASCs) have recently emerged as a more viable source for clinical applications, compared to bone-marrow mesenchymal stromal cells (BM-MSCs) because of their abundance and easy access. In this study we evaluated the regenerative potency of ASCs compared to BM-MSCs. Furthermore, we compared the dielectric and electro-kinetic properties of both types of cells using a novel Dielectrophoresis (DEP) microfluidic platform based on a printed circuit board (PCB) technology. Our data show that ASCs were more effective than BM-MSCs in promoting neovascularization in an animal model of hind-limb ischemia. When compared to BM-MSCs, ASCs displayed higher resistance to hypoxia-induced apoptosis, and to oxidative stress-induced senescence, and showed more potent proangiogenic activity. mRNA expression analysis showed that ASCs had a higher expression of Oct4 and VEGF than BM-MSCs. Furthermore, ASCs showed a remarkably higher telomerase activity. Analysis of the electro-kinetic properties showed that ASCs displayed different traveling wave velocity and rotational speed compared to BM-MSCs. Interestingly, ASCs seem to develop an adaptive response when exposed to repeated electric field stimulation. These data provide new insights into the physiology of ASCs, and evidence to their potential superior potency compared to marrow MSCs as a source of stem cells.

No MeSH data available.


Related in: MedlinePlus