In vivo 3 dimensional cartilage tissue engineering by adipose derived stem cells

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The species of adipose derived stem cells was changed from human to GFP transgenic mice in this project because human specimen was relatively difficult to obtain. Accordingly, we firstly identified that similar population of human adipose derived stem cells (ASCs) can be isolated from GFP transgenic mice.After the proliferation of ASC in control medium, they were induced into chondrogenic differentiation in the induction medium using micromass culture technique for 2 week. Subsequently, ASCs were successfully differentiated into spheroid-shape cartilage of 1 to 2mm in diameter. This culture technique was also applied to regenerate relatively big size of cartilage which is 8 to 10mm in diameter without any scaffolds. When these structures were implanted in vivo into the vascular-rich space such as subfascia or intramuscular region, they were found to keep in shape and size both macroscopically and histologically.Next, we investigated whether artificial polypeptide (PuraMatrix^<TM>) could be suitable scaffold for chondrogenic regeneration using ASCs. 1% PuraMatrix^<TM> with 3x10^6 ASCs were incubated in the chondrogenic media and observed up to 8 weeks. After 2 weeks of incubation, these mixtures exhibited the production of the chondrogenic matrices which was confirmed by Alcian blue staining. In addition, the phenotype was still identical after 8 weeks of incubation.