[Preliminary study on tissue-engineered cartilage with human dermal fibroblasts co-cultured with porcine chondrocytes in vitro].
OBJECTIVE
To explore the feasibility of constructing tissue-engineered cartilage with human dermal fibroblasts (HDFs) in vitro.


METHODS
Porcine articular chondrocytes and HDFs were isolated and in vitro expanded respectively. Then they were mixed at the ratio of 1:1 (chondrocytes: fibroblasts) . The mixed cells were seeded onto polyglycolic acid (PGA) scaffold at the ultimate concentration of 5.0 x 10(7)/ml as co-culture group. Chondrocytes and HDFs at the same ultimate concentration were seeded respectively onto the scaffold as chondrocyte group ( positive control group) and fibroblast group ( negative control group). The specimens were collected after in vitro culture for 8 weeks. Gross observation, histology and immunohistochemistry were used to evaluate the results.


RESULTS
In chondrocyte group, the cell-scaffold constructs could maintain the original size and shape during in vitro culture. The new formed cartilage-like tissue had typical histological structure and extracellular matrix staining similar to normal cartilage. In co-culture group the constructs shrunk slightly at 8 weeks, cartilage-like tissue formed and GAG could be detected for strong expression by Safranin O staining. Furthermore, using the specific identification, a few HDFs derived cells were found to form lacuna structure at the peripheral area of cartilage-like tissue. In fibroblast group, the constructs deformed and shrunk gradually without mature cartilage lacuna in histology.


CONCLUSIONS
The 3D-co-culture system can effectively induce the differentiation of HDFs to chondrocytes. The tissue-engineered cartilage can be constructed in vitro with the 3D-co-culture system.
Liu
Xia
X
Plastic Surgery Hospital, Chinese Academy of Medical Science, Beijing 100144, China.
Zhou
Guang-dong
GD
Liu
Wei
W
Cao
Yi-lin
YL
D000818:Animals; D002356:Cartilage / Q000166:cytology*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology; D018920:Coculture Techniques; D020405:Dermis / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds