Osteogenic and adipogenic differentiation of rabbit adipose-derived mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2014.37.019

Abstract

Abstract:

BACKGROUND: Adipose-derived mesenchymal stem cells have the ability to self-renew and have pluripotent potential under specific conditions in vitro, which have broad application prospects in clinical practice. However, isolation and culture of adipose-derived mesenchymal stem cells still appear to have many difficulties and shortcomings.
OBJECTIVE: To isolate and culture rabbit adipose-derived mesenchymal stem cells in vitro in order to study their morphology, cell surface markers and biological properties as well as to investigate the osteogenic and adipogenic potentials of adipose-derived mesenchymal stem cells in vitro.
METHODS: Primary adipose-derived mesenchymal stem cells were isolated from the subcutaneous adipose tissue of posterior cervical region from New Zealand white rabbits and digested by 0.1% collagenase I. The cells were passaged and amplified by the trypsin digestion. The passage 4 adipose-derived mesenchymal stem cells were induced to differentiate after exposure to adipogenic or osteogenic medium. The oil red O staining, alkaline phosphatase and alizarin red staining were used to detect the results. The cell viability was detected by the cell counting kit 8 method to drawn the growth curve. Cell surface markers were examined using flow cytometry.
RESULTS AND CONCLUSION: The adipose-derived mesenchymal stem cells isolated from the subcutaneous adipose tissue of rabbits exhibited a fusiform adherent growth in a vortex pattern, and had a strong capability of proliferation that could be passaged stably to the 10th generation. Flow cytometry results showed that the cells highly expressed CD29, CD90, CD44, but lowly expressed CD45 and CD34. After adipogenic induction, the adipose-derived mesenchymal stem cells were positive for oil red O staining; after osteogenic induction, the cells were both positive for alkaline phosphatase and alizarin red staining. These findings suggest that the adipose-derived mesenchymal stem cells were successfully isolated and cultured from the subcutaneous adipose tissue of rabbits, and these cells are pluripotent with the potential to differentiate into adipocytes and osteoblasts, which are expected to be ideal seed cells for bone tissue engineering.

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

全文链接：

Key words: subcutaneous fat, mesenchymal stem cells, rabbits, adipogenesis

CLC Number:

R394.2

Lee Soo-min, Wu Zi-zheng, Wang Zhe, Li Zhi, Zhang Jian . Osteogenic and adipogenic differentiation of rabbit adipose-derived mesenchymal stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(37): 6008-6013.

Figures/Tables 3

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