Adipose-derived stem cells: isolation, culture and differentiation into endothelial progenitor cells

doi:10.3969/j.issn.2095-4344.2015.32.018

Abstract

Abstract:

BACKGROUND: Adipose-derived stem cells are regarded as the potential seed cells for tissue engineering. Collagenase digestion is used to isolate adipose-derived stem cells from fat pads currently. However, there are some problems, such as cumbersome operation and high cost.
OBJECTIVE: To study the basic biological characteristics of adipose-derived stem cells by tissue explants culture and to explore the differentiation potential into osteoblasts, adipocytes and endothelial progenitor cells in vitro.
METHODS: Adipose-derived stem cells were isolated by tissue explants technique from the bilateral groin fat pads of rats under aseptic conditions, and cultured in vitro. Cell counting kit-8 was used to detect the proliferative activity, and flow cytometry was employed to analyze the expression of cell surface markers. Passage 4 adipose-derived stem cells were cultured in osteogenic medium, adipogenic medium and endothelial progenitor cell medium for 2-3 weeks, and then the cells were identified.
RESULTS AND CONCLUSION: Adipose-derived stem cells that were isolated by tissue explants culture were easily cultured, and after subculture, cells were mainly spindle-shaped and grew in clone-like manner with swirling arrangement. Cells that experienced repeated subcultures still kept stronger proliferative ability and the cell growth curve was shaped as a parabola. Immunochemical staining analysis revealed that adipose-derived
stem cells were positive for CD44, CD90 and CD29, but negative for CD31, CD45. After adipogenic/osteogenic induction, the cells were respectively positive for oil red O staining and alizarin red staining. Induced endothelial progenitor cells were identified with CD34 and the ability to uptake Dil-ac-LDL and FITC-UEA. These findings indicate using the using tissue explants culture, high-purity adipose-derived stem cells easy to proliferate can be harvested, highly express stem cells-related antigens, and have the ability to differentiate into osteoblasts, adipocytes and endothelial progenitor cells, which meet the needs of seed cells in tissue engineering research.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Adipose Tissue, Cell Differentiation, Endothelial Cells

CLC Number:

R394.2

Liu Zi-qi, Sun Tong-wen, Wan You-dong, Yao Rui, Kan Quan-cheng. Adipose-derived stem cells: isolation, culture and differentiation into endothelial progenitor cells [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(32): 5182-5187.

Figures/Tables 3

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