Co-culture of adipose derived stem cells and chondrocytes

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Abstract

BACKGROUND:

Study shows that bone marrow mesenchymal stem cells can be induced into chondrocytes in the microenvironment of articular cartilage.

OBJECTIVE:

To verify the feasibility of inducing adipose derived stem cells (ADSCs) into chondrocytes so as to provide a novel approach for articular cartilage tissue engineering.

DESIGN, TIME AND SETTING:

A randomized cell trial was performed at the State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University from September 2007 to July 2008.

MATERIALS:

Adult New Zealand white rabbits of clean grade, weighing 1.5 to 2.0 kg, of either gender.

METHODS:

Primary ADSCs and aricular chondrocytes from New Zealand white rabbits were isolated and then seeded on 6-pore plates and millicell culture plate inserts separately. After 2 weeks of co-culture, trypsinization was terminated and ADSCs were detected with oil red O staining method, alkaline phosphatase detection method and toluidine blue staining method respectively.

MAIN OUTCOME MEASURES:

The cytomorphology of ADSCs was observed under inverted microscope before and after co-culture respectively. After co-culture, the proteoglycan expression of ADSCs was detected with toluidine blue staining method and the collagen type II expression with immunocytochemistry detection method. The Genetic transcription level of aggrecan and collagens type II was determined with reverse transcriptase-polymerase chain reaction.

RESULTS:

All of the three detection method (namely, oil red O staining method, alkaline phosphatase detection method and toluidine blue staining method) proved that ADSCs had the multi-directional differentiation ability and were mesenchymal stem cells derived from adiposes. After 1 week of co-culture, parts of ADSCs were rounded morphologically. The genetic transcription level and the expression of both aggrecan and collagens type II increased after 2 weeks of co-culture.

CONCLUSION:

ADSCs are able to be induced into cartilage-like cells when being co-cultured with chondrocytes.

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