Adipose-derived Stem Cells: Isolation, Culture And Differentiation Into Endothelial Progenitor Cells

Background and Objective Orthopedics hairdressing industry has been rapidly developed in recent years, It can obtained a lot of adipose derived stem cells from a simple fat suction to provide sufficient cells for experimental and clinical studies, what’s more, the adipose derived stem cells have not been involved in the ethical issues. Therefore, Adipose derived stem cells,as adult stem cells, can instead of the embryonic stem cells to become the seed cell of tissue engineering.However, The traditional method of obtaining adipose derived stem cells is the trypsin digestion method considered to be a time-consuming and inefficient method.An urgent need to get a fast and efficient method to separate and culture the adipose derived stem cells, to meet a lot of cells in demand for basic and applied research,Tissue block culture method can not only ensure the adipose stem cells with strong proliferation ability, and keep low levels of cellular senescence in long-term culture in vitro. It is an effective method to obtain a large number of high quality adipose derived stem cells.MethodsAnaesthetize the SD rats and cut adipose tissue from the inguinal of rats under a sterile condition to get adipose derived stem cells by using the tissue culture method,detect the cell proliferation activity of adipose derived stem cells by CCK-8, test the adipose derived stem cell surface marker expression of markers by flow cytometry.Inducing adipose derived stem cells differentiate into bone cells, fat cells and endothelial progenitor cells, and test the surface markers of these cells.Resultswe can obtain adipose derived stem cells in vitro by the tissue culture method,the cells are amplified easily, the shape of the most of cells are long fusiform, the growth of cells are quickly and the morphology and characteristics are stable, the cells can be passaged for the dozens of generations, as also maintain a strong proliferation, the growth curve of the cells show a parabola shape; flow cytometry is used to detect the cell surface markers, show that positive expression on CD44, CD90 and CD29, negative expression on the other ones; And the cells could be induced to adipocytes, which are positive in Oil red O stains; as well to osteoblast cells, which are positive in alizarin red staining; the adipose derived stem cells in vitro can be induced to endothelial progenitor cells, cell surface markers CD31, CD34, CD133,KDR can be detected positive, and Di I-ac-LDL and FITC-UEA-1 double fluorescent staining are tested positive.ConclusionIt can be obtained adipose derived stem cells by tissue culture method successfully, the cells with high purity are easy to be cultured, and the surface markers are expressed highly, as the same time, they are of the multi-directional differentiation ability to induce the osteoblasts, adipocytes and endothelial progenitor cells, to demand a full foot tissue engineering seed cells definitely.