Multi-directional induced differentiation of rabbit adipose-derived stem cells

doi:10.3969/j.issn.2095-4344.2012.49.008

Abstract

Abstract:

BACKGROUND: Studies have shown that adipose-derived stem cells can be induced to differentiate into osteoblasts, chondrocytes, myocardial cells, neurons, epithelial cells and hepatocytes under certain conditions.
OBJECTIVE: To investigate the isolation and culture method and the basic biological characteristics of adipose-derived stem cells in order to identify its cell phenotype.
METHODS: Adipose tissue was collected from the neck and back of adult Japanese white rabbits, and the primary adipose-derived stem cells were obtained and cultured with type Ⅰ collagenase and when the cells grow to cover about 80% of the space, the growth situation and morphological characteristics of adipose-derived stem cells were observed with inverted microscope every day, and the growth curve was drawn; cell surface markers were checked by flow cytometry; the adipose-derived stem cells were induced to osteogenesis and adipogenesis respectively, the osteogenic differentiation potential and adipogenic differentiation potential were assessed through the alkaline phosphatase, alizarin red, VonKossa (calcium nodules) staining and oil red O staining of adipose-derived stem cells, control group was non-induced.
RESULTS AND CONCLUSION: Adipose-derived stem cells cultured in vitro exhibited a spindle-shaped appearance and actively proliferated. After subculture, adipose-derived stem cells were strongly proliferated and their growth curve was “S” shaped. Passages 3 and 6 adipose-derived stem cells highly expressed CD29 and CD44 as detected by flow cytometry, with a positive rate of more than 90% and lowly expressed CD34 and CD45 with a positive rate of less than 5%; and the expression of CD29 and CD44 was gradually increased while the expression of CD34 and CD45 was gradually decreased with the passage increased. Alkaline phosphatase, alizarin red and von Kossa staining were positive in the osteogenic-induction group and oil red O staining was positive in the adipogenic-induction group, whilethese staining were negative in the control group. Rabbit adipose-derived stem cells could be easily isolated and cultured, and stably proliferated, and they expressed mesenchymal stem cells-related phenotype and could be induced to differentiate into osteoblasts and adipocytes under certain conditions.

CLC Number:

R394.2

Wang Qing-fu,Chen Zhuang-hong, Cai Xian-hua, Xie Yong-hui, Diao Bo, Liu Qin. Multi-directional induced differentiation of rabbit adipose-derived stem cells[J]. Chinese Journal of Tissue Engineering Research, 2012, 16(49): 9162-9167.

Figures/Tables 5

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