Normal features of tissue-engineered auricular cartilage by flow cytometry and histology: patient safety.
BACKGROUND
Cytokinetic abnormalities in DNA content, such as aneuploidy, haploidy, and tetraploidy, have been found to occur in human cartilaginous tumors. The high number of chondrocytes needed for tissue-engineered cartilaginous implants requires the cells to be passaged repeatedly. The theoretical risk of changes in the normal diploid state of these cells during their growth in vitro and after generation of tissue-engineered cartilage in vivo is not known. Materials and methods Auricular chondrocytes were obtained from 6 patients and cultured in vitro. Chondrocyte number was increased by repeated passaging. The passaged cells were implanted in nude mice for 8 weeks to generate tissue-engineered cartilage. Fresh control chondrocytes along with the passaged cells and cells obtained from the tissue-engineered constructs were collected and compared for DNA content by flow cytometry.


RESULTS
Flow cytometry demonstrated 100% diploidy with no evidence of aneuploidy, haploidy, or tetraploidy in all groups of cells. Histology of the tissue-engineered cartilage also showed no evidence of cellular atypia.


CONCLUSIONS
The number of human auricular chondrocytes can be increased by repeated passaging and passaged chondrocytes can be safely used for implantation to generate tissue-engineered constructs without a change in the normal diploid state of the cells. Histology of the cartilage generated showed normal features without atypia.
Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, Boston, MA 02114, USA.
Syed H
SH
Kamil
Marcia
M
Woda
Lawrence J
LJ
Bonassar
Yuri W
YW
Novitsky
Charles A
CA
Vacanti
Roland D
RD
Eavey
Martin P
MP
Vacanti
D000782:Aneuploidy; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D004425:Ear Cartilage / Q000166:cytology*; D004428:Ear Neoplasms / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D005818:Genetic Engineering / Q000009:adverse effects / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D012449:Safety