Pelleted cells were suspended with 5 mmol/l EDTA/PBS and layered over an equal volume of 1 . 083 g/ml Histopaque 1083 solution (Sigma-Aldrich Japan K. K., Tokyo, Japan, http://www.sigmaaldrich.com). tissue-derived AdSCs. Immunocytochemistry showed that CA-derived AdSCs had the highest appearance rates of markers for endothelial cells, vascular smooth muscle cells, and cardiomyocytes among the AdSCs. Systemic transfusion of CA-derived AdSCs exhibited the highest cardiac functional recovery after myocardial infarction and the high frequency of the recruitment to ischemic myocardium. Moreover, long-term follow-up of the recruited CA-derived AdSCs frequently expressed cardiovascular cell markers compared with the other adipose tissue-derived AdSCs. Cardiac adipose tissue could be an ideal source for isolation of therapeutically effective AdSCs for cardiac regeneration in ischemic heart diseases. == Significance == The present study found that cardiac adipose-derived stem cells have a high potential to differentiate into cardiovascular lineage cells (i. e., cardiomyocytes, endothelial cells, and vascular smooth muscle cells) compared with stem cells derived from other adipose tissue such as subcutaneous, visceral, and subscapular adipose tissue. Notably, only a small number of supracardiac adipose-derived stem cells that were systemically transplanted sufficiently Fatostatin improved cardiac functional recovery after myocardial infarction, differentiating into cardiovascular cells in the ischemic myocardium. These findings suggest a new autologous stem cell therapy for patients with myocardial ischemia, especially those with secondary myocardial ischemia after cardiovascular open chest surgery. == Introduction == Heart failure resulting from ischemic heart disease is one of the leading causes of hospitalization and reduced quality Fatostatin of life for patients worldwide [1]. Although advances in the treatment of patients with myocardial infarction (MI), such as the development of devices for catheterization, including drug-eluting stents, have significantly improved the outcomes, a need for novel therapies remains to reduce mortality further and prevent the deleterious consequences that result from myocardial damage. At present, the only effective treatment for advanced heart failure that can fully restore cardiac function is heart transplantation. However , only a limited number of patients will be able to receive the therapy owing to the difficulty of finding appropriate donors and/or legal restrictions in each country. Recently, numerous investigators have evaluated the application of stem/progenitor cells for Rabbit Polyclonal to ARMCX2 the treatment of ischemic heart diseases, including MI, in both animal experiments and human clinical trials, and favorable outcomes have been demonstrated. The major stem/progenitor cell sources for human autologous cell transplantation therapy have been reported to be peripheral blood [24] and bone marrow [47] Fatostatin in ongoing clinical trials for ischemic heart diseases. However , each of the currently used adult stem cell reservoirs has practical limitations that obstruct their widespread use in treating human disease owing to the requirement of invasive harvesting procedures or ex vivo manipulation/expansion techniques. Adipose tissue consists of mature adipocytes and a mononuclear cell fraction termed stromal vascular fraction (SVF). The SVF is a diverse mixture of cells, including endothelial cells (ECs), vascular smooth muscle cells (VSMCs), blood cells, and mesenchymal stem cells (MSCs), that is identical to adipose tissue-derived stem cells (AdSCs). AdSCs have phenotypic and functional properties (i. e., multilineage differentiation potential) similar to those of bone marrow-derived MSCs. Of particular relevance, AdSCs have been reported to differentiate, not only into adipocytes [8], osteoblasts [8, 9], chondrocytes [10, 11], pancreatic -cells [12], hepatocytes [13], neural Fatostatin cells [14, Fatostatin 15], and myocytes [8], but also into cardiovascular lineage cells, such as cardiomyocytes (CMs) [16], ECs [9, 17], and VSMCs [18]. Most importantly for their clinical application, AdSC-enriched SVFs can be isolated in large quantities by minimally invasive liposuction, with a significantly higher yield of progenitor cells per volume compared with bone marrow. The SVF of adipose tissue has the potential to improve cardiac function after MI by several mechanisms: delivery of cardiovascular cell replacement, salvage of host cardiomyocytes through antiapoptotic mechanisms, or stimulation of angiogenesis. Similar to bone marrow-derived MSCs, AdSC-enriched SVFs secrete a number of paracrine factors that are proangiogenic [19], anti-inflammatory [20, 21], and chemoattractive for stem/progenitor cells [19] and demonstrate their beneficial effects on jeopardized myocardium by ischemic insult. Early clinical trials of autologous AdSC-enriched SVF transplantation therapy for MI patients are now ongoing in Europe and have shown significant improvement in cardiac function [22]. However , the favorable effect of transplanted SVF has been attributed to the paracrine effect rather than a direct contribution of AdSCs to tissue regeneration via transdifferentiation into cardiovascular lineage cells, as we reported previously [19]. To use AdSCs as a tool for myocardial tissue regeneration therapy, the frequency of AdSC transdifferentiation into cardiovascular cells should be increased either by manipulation of the inducible culture method or selection of appropriate adipose.