Isolation, culture and osteogenic differentiation of adipose-derived stem cells from the abdominal cavity of rats

doi:10.3969/j.issn.2095-4344.2013.23.008

Abstract

Abstract:

BACKGROUND: Adipose-derived stem cells could be obtained easily with rapid proliferation and multi-directional differentiation potential, so they are promising to be seed cells of tissue engineering instead of bone marrow stromal stem cells. However, the isolation and culture of adipose-derived stem cells still remain lots of difficulties.
OBJECTIVE: To optimize the methods of isolation and culture of adipose-derived stem cells, and identify the ability of osteogenic differentiation.
METHODS: The adipose tissues were obtained from the kidney, peri-uterus and abdominal cavity of one adult female rat weighing 200 g under sterile conditions. Adipose-derived stem cells were isolated by the method of collagenase digestion for several times and cultured with low-glucose medium. The morphologic changes and proliferation of adipose-derived stem cells were observed under inverted microscope. The passage 3 adipose-derived stem cells were cultured in osteogenic medium and then identified with alkaline phosphatase staining and alizarin red staining.
RESULTS AND CONCLUSION: Adipose-derived stem cells were mainly spindle-shaped and proliferated quickly with whirlpool-shaped arrangement. After osteogenic induction for 10 days and 28 days, the results of alkaline phosphatase staining and alizarin red staining were both positive. Experimental findings indicate that, adipose-derived stem cells isolated from abdominal cavity of rat by the method of collagenase digestion for several times can easily be cultured and passaged in vitro, and allow stable passaging and differentiation into osteoblasts under certain induction conditions. The isolation and culture methods for the adipose-derived stem cells may offer more and better seed cells for tissue engineering.

Key words: stem cells, adipose-derived stem cells, seed cells, isolation and culture, osteogenic differentiation, induction, abdominal cavity, mineralization nodules, alkaline phosphatase, orthopedic surgery, tissue engineering, National Natural Science Foundation of China, stem cell photographs-containing paper

CLC Number:

R394.2

Li Ling-hui, Ding Dao-fang, Gong Hao, Du Guo-qing, Song Yi, Deng Zhen, Zhan Hong-sheng. Isolation, culture and osteogenic differentiation of adipose-derived stem cells from the abdominal cavity of rats[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(23): 4232-4239.

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