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Abstract

BACKGROUND:

A great amount of mesenchymal stem cells can be successfully derived from fat tissue and induced to differentiate into osteoblasts, chondrocytes, adipocytes and myocardial cells.

OBJECTIVE:

To establish the method of isolating, culturing and osteogenic differentiation of adipose-derived stem cells in vitro, and to explore the potential of adipose-derived stem cells as seed cells for bone tissue engineering.

METHODS:

Collagenase enzymatic digestion was used to isolate adipose-derived stem cells from human fat tissue which were then cultured in vitro. Flow cytometry was used to detect cell surface markers. Cell counting kit-8 assay was performed to examine cell viability. Adipose-derived stem cells were induced by osteogenic induced reagent to differentiate into bone cells. In addition, we also performed BCIP/NBT method to detect alkaline phosphatase activity. Alizarin red staining was used to detect the formation of calcium node. RT-PCR was performed to examine alkaline phosphatase and osteopontin expression.

RESULTS AND CONCLUSION:

We successfully obtained adipose-derived stem cells from fat aspirated liquid. Adipose-derived stem cells obtained could passage stably and proliferate highly. Flow cytometry data showed the expression of stem cell surface markers. Adipose-derived stem cells showed typical osteoblast morphology after

RESULTS AND CONCLUSION:

osteogenic differentiation. Alkaline phosphatase staining was positive and alizarin red staining displayed the formation of calcium node. Furthermore, we found that alkaline phosphatase and osteopontin mRNA was expressed after differentiation 0, 3, 7, 14, 21, 28 days. These findings indicate that adipose-derived stem cells can be obtained from fat tissue through enzymatic digestion, differentiate towards bone cells, and express alkaline phosphatase and osteopontin, which can become potential seed cells for bone tissue engineering.

RESULTS AND CONCLUSION:

Funding: Key Provincial Talents Program of Jiangsu Province, No. RC2011115

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