Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo
Section snippets
Methods
Materials. Collagenase II was obtained from Sigma. Dispase was purchased from Gibco (Gibco Life Technologies, Paisley, UK). For the fluorescence-activated cell sorter (FACS) analyses, and the anti-human CD29, CD31, CD34, CD44, CD45, CD105, CD106, CD184, HLA-ABC, Flk1 and the control antibodies were from BD Bioscience. For the endocytosis function analysis, DiI-labeled acetylated LDL was provided by Molecular Probes.
Preparation of human ADAS cells from the adult human fat. Human raw
Morphology and phenotypes of cultured ADAS cells
The adherent cultured ADAS cells assumed a fibroblast-like morphology when observed under a light microscope. The morphology was maintained through repeated subcultures under non-stimulating conditions. At the log phase, the doubling time was about 22 h. To characterize the phenotypes of adherent adipose-derived cells, flow cytometry was performed. The results showed that these cultured cells were positive for CD29, CD44, CD105, CD166, Flk1, and HLA-ABC. In addition, no expression of the
Discussion
The vascular endothelium represents a dynamic border between circulating blood and the surrounding tissue. In healthy subjects, a low basal level of endothelial turnover (1–3/ml blood) has been described [18]. During acute injury of the vascular endothelium, endothelial reconstruction can occur by migration and proliferation of surrounding mature ECs. However, mature ECs are terminally differentiated cells with a low proliferative potential, and their capacity to substitute damaged endothelium
Acknowledgments
This work was supported by grants from the “863 Projects” of Ministry of Science and Technology of PR China (No 2002AA205061); from China Medical Board of New York, Inc.: Stem Cell Biology, Engineering (Grant #01-748); from National Natural Science Foundation of China (No. 30070284); from National Key Project for Basic Research of China (No. 001CB5099); and from Beijing Ministry of Science and Technology (No. 2002-489); R.C.H. Zhao is a Cheung Kong Scholar in PR China.
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