Research of co-culture of adipose derived stem cells and chondrocytes in different culture conditions / 中国矫形外科杂志

ABSTRACT

[Objective]To investigate the feasibility and optimal condition of in vitro chondrogenesis by co-culture of adipose derived stem cell(ADSC)and chondrocytes in pathologic state so as to provide a new clinical approach for repairing damaged articular cartilage.[Method]ADSC and aricular chondrocytes from osteoarthritis model in adult New Zealand white rabbits were in vitro expanded and then seeded on plate and plug-in type millicell dish respectively.The co-culture cells were cultured in different conditions including different serum concentration(10% FBS and 2% FBS)and dimensions(fibrin scaffold and monolayer).The culture medium was changed every 3 days.The shape of ADSC before and after co-culture was observed by inverted microscope and transmission electron microscope.The expression of aggrecan and type Ⅱ collagen genes in ADSC were studied by toluidine blue staining and immunohistochemistry after in vitro co-culture for 14 days.And the transcription of aggrecan and type Ⅱ collagen genes in ADSC were studied by RT-PCR.[Result]The shape of ADSC was fibroblast-like cells morphologically.And they became round at 7 days after in vitro co-culture.At 14 days after co-culture with chondrocytes,ADSC was changed to chondrocyte-like cells morphologically and increased immunostaining particles of type Ⅱ collagen and enhanced toluidine blue staining.And the transcription of type Ⅱ collagen and aggrecan genes was also increased,especially in the ADSC cultured in fibrin scaffold with 10% FBS.[Conclusion]Chondrocytes in pathologic state may provide chondrogenic microenvironment to induce chondrogenic differentiation of ADSC in vitro,which will be enhanced by high serum concentration and three-dimensional culture.