title: Tissue-engineered human auricular cartilage demonstrates euploidy by flow cytometry.
abstract: Transforming growth factor-beta (TGF-beta) and basic fibroblast growth factor (bFGF) are known to stimulate the rate of chondrocyte proliferation. The theoretical risk of malignant transformation associated with growth factor stimulation of chondrocytes should be addressed; aneuploidy has been found to occur in human cartilaginous tumors. In this study, chondrocytes were obtained from six human auricles and cultured in vitro for 6 weeks in the presence or absence of TGF-beta and bFGF. Cells were analyzed for DNA at 3-, 4-, 5-, and 6-week intervals by flow cytometry (FACScan), which demonstrated no evidence of aneuploidy. A persistent increase in S-phase was noted in cells cultured only with TGF-beta. Cells were implanted in athymic mice, and after 8 weeks of implantation, the cartilage constructs formed were examined histologically. The tissue-engineered cartilage cultured originally in bFGF most resembled normal, native cartilage. Specimens cultured in TGF-beta produced suboptimal cartilage morphology. Flow cytometry shows no evidence of aneuploidy, with chondrocytes maintaining their normal diploid state. Further studies incorporating additional methods of analysis need to be done.
authors.affiliation: Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, and the Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: S H
authors.initials: SH
authors.lastname: Kamil
authors.forename: B S
authors.initials: BS
authors.lastname: Aminuddin
authors.forename: L J
authors.initials: LJ
authors.lastname: Bonassar
authors.forename: C A Arevalo
authors.initials: CA
authors.lastname: Silva
authors.forename: Y
authors.initials: Y
authors.lastname: Weng
authors.forename: Marcia
authors.initials: M
authors.lastname: Woda
authors.forename: C A
authors.initials: CA
authors.lastname: Vacanti
authors.forename: R D
authors.initials: RD
authors.lastname: Eavey
authors.forename: M P
authors.initials: MP
authors.lastname: Vacanti
mesh_terms: D000293:Adolescent; D000818:Animals; D002356:Cartilage / Q000166:cytology* / Q000637:transplantation / Q000648:ultrastructure; D002675:Child, Preschool; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D011003:Ploidies; D023822:Tissue Engineering; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D019737:Transplants
chemical_list: D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2