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Tissue-engineered human auricular cartilage demonstrates euploidy by flow cytometry.
S H Kamil1, B S Aminuddin, L J Bonassar
1Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, and the Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts, USA.
Tissue Engineering
|March 12, 2002
Summary
Transforming growth factor-beta (TGF-beta) and basic fibroblast growth factor (bFGF) did not induce aneuploidy in cultured human chondrocytes. Tissue-engineered cartilage grown with bFGF showed the best resemblance to native cartilage.
Area of Science:
- Tissue engineering
- Cell biology
- Biomaterials science
Background:
- Growth factors like TGF-beta and bFGF stimulate chondrocyte proliferation.
- Malignant transformation and aneuploidy are theoretical risks in growth factor-stimulated chondrocytes.
- Aneuploidy is observed in human cartilaginous tumors.
Purpose of the Study:
- To investigate the potential for aneuploidy in human chondrocytes cultured with TGF-beta and bFGF.
- To evaluate the histological quality of tissue-engineered cartilage formed under these conditions.
Main Methods:
- Human auricular chondrocytes were cultured in vitro for 6 weeks with or without TGF-beta and bFGF.
- DNA analysis for aneuploidy was performed using flow cytometry at multiple time points.
- Engineered cartilage constructs were implanted in athymic mice and examined histologically after 8 weeks.
Main Results:
- Flow cytometry revealed no evidence of aneuploidy; chondrocytes maintained a diploid state.
- Cells cultured with TGF-beta showed a persistent increase in S-phase.
- Tissue-engineered cartilage cultured with bFGF most closely resembled native cartilage histologically.
- TGF-beta resulted in suboptimal cartilage morphology.
Conclusions:
- TGF-beta and bFGF stimulation of human chondrocytes in vitro does not lead to aneuploidy.
- bFGF promotes the formation of tissue-engineered cartilage with morphology similar to native cartilage.
- Further analysis is recommended to fully assess chondrocyte behavior and safety.