Three-dimensional culture method induces the chondrogenic differentiation of human adipose-derived mesenchymal stem cell microspheres

doi:10.3969/j.issn.2095-4344.2015.01.005

Abstract：BACKGROUND:Adipose-derived mesenchymal stem cels are more appropriate as seed cels for cartilage induction in patients who have unsatisfied repair after articular cartilage injuries, but how to induce the cartilage function need to be studied. OBJECTIVE:To induce the chondrogenic differentiation of human adipose-derived mesenchymal stem cel microspheres using three-dimensional culture system. METHODS:Adipose tissues were extracted after liposuction under sterile, to isolate and culture human adipose-derived mesenchymal stem cels. The third passage cels were analyzed by flow cytometry and identified by osteogenic induction and adipogenic induction, and meanwhile, cels were cultured under three-dimensional culture system for chondrogenic differentiation. Then, cels were observed by Alcian blue staining for identification of glycosaminoglycan synthesis as wel as hematoxylin-eosin staining for histological analysis. Expression of colagen type II was detected by immunofluorescence method. Cartilage stiffness was determined by mass measurement. RESULTS AND CONCLUSION:Isolated human adipose-derived mesenchymal stem cels highly expressed CD105, CD44, CD29, but showed low expressions of CD45, CD34. After osteogenic and adipogenic induction, cels were positive for alizarin red staining and oil red O staining. Under the three-dimensional culture, chondrocytes could greatly express glycosaminoglycans and colagen type II. The results suggest that under the three-dimensional culture, human adipose-derived mesenchymal cels differentiate into chondrocytes and have the characteristics of chondrocytes.