Transplantation of freshly isolated adipose tissue-derived regenerative cells enhances angiogenesis in a murine model of hind limb ischemia

Abstract

Therapeutic angiogenesis has emerged as one of the most promising therapies for severe ischemic cardiovascular diseases with no optional therapy. Several investigators have reported that transplantation of cultured adipose-derived regenerative cells (cADRCs) to ischemic tissues promotes neovascularization and blood perfusion recovery; however, cell therapy using cultured cells has several restrictions. To resolve this problem, the angiogenic capacity of freshly isolated ADRCs (fADRCs) obtained from Lewis rats was compared with cADRCs, both in vivo and in vitro. Flow cytometric analysis showed that fADRCs contained several cell types such as endothelial progenitor cells and endothelial cells; however, these cells were present in a very small proportion in cADRCs. Transplantation of fADRCs in mice significantly improved blood perfusion, capillary density, and production of several angiogenic factors in transplanted ischemic limbs compared with a saline-injected group, whereas these effects were not observed in the cADRCs-injected group. fADRCs also showed significantly higher expression levels of angiogenic factors than cADRCs in the in vitro study. Furthermore, fADRC stimulated tube formation more remarkably than cADRC in an in vitro tube formation assay. These results suggested that fADRCs have an effective angiogenic capacity, and they would be more valuable as a source for cell-based therapeutic angiogenesis than cADRCs or other stem/progenitor cells.

<b>VEGF secretion by adipose tissue-derived regenerative cells is impaired under hyperglycemic conditions via glucose transporter activation and ROS incr</b><b>ease </b>

“…ADRCs expressing GLUT1, 3 and 4 uptake excess markers such as CD13, CD49, CD44, CD90, and CD105 (18). We previously found that although freshly isolated ADRCs included CD31, CD34, and CD45 positive cells, CD90 positive cells became predominant during culture, which indicated proliferation of only mesenchymal cells during culture (5). Under hyperglycemic conditions the predominance of CD90 positive cells was not affected, indicating that hyperglycemia does not influence the subpopulation of ADRCs.…”

<b>Adipose stem cell sheets improved cardiac function in the rat myocardial infarction, but did not alter cardiac contractile responses to β-adrenergic stim</b><b>ulation </b>

“…ASCs were enzymatically isolated from the inguinal subcutaneous fat tissue of rats and cultured as previously described (3). Skeletal myoblasts were isolated from the skeletal muscle of the thigh as described previously (17).…”

“…Interestingly, ASC sheets expressed significantly higher mRNA and protein levels of HGF, VEGF, and bFGF than myoblast cell sheets under hypoxic condition, associated with the less prevalence of cell apoptosis in ASC sheets than myoblast cell sheets. bFGF also augments neovascularization and inhibits cellular apoptosis (3,21). Since preventing apoptosis of transplanted cells leads to improvement of Fig.…”

“…mRNA levels were expressed relative to the levels of β-actin. The primers were designed to span their genomic regions described elsewhere (3). Same condition of RT-PCR was used to quantify mRNA of heart tissues as described in in vivo experiments.…”

Angiogenic Synergistic Effect of Adipose-Derived Stromal Cell Spheroids with Low-Level Light Therapy in a Model of Acute Skin Flap Ischemia

“…The advantage of ASCs includes their relative abundance, rapid expansion, multipotency and low immunogenicity as compared with other mesenchymal stromal cells [Gimble, 2003;Huang et al, 2004]. Transplanting ASCs induces neovascularization and improves blood flow to ischemic tissue in animal models [Sumi et al, 2007;Harada et al, 2013]. However, less research has been conducted on transplanted xenogeneic ASCs to determine whether endothelial differentiation is evident [Gong et al, 2014].…”