title: Quantitative tracking of passage and 3D culture effects on chondrocyte and fibrochondrocyte gene expression.
abstract: Success in cartilage and fibrocartilage tissue engineering relies heavily on using an appropriate cell source. Many different cell sources have been identified, including primary and stem cells, along with experimental strategies to obtain the required number of cells or to induce chondrogenesis. However, no definitive method exists to quantitatively evaluate the similarity of the resulting cell phenotypes to those of the native cells between candidate strategies. In this study, we develop an integrative approach to enable such evaluations by deriving, from gene expression profiles, two quantitative metrics representing the nearest location within the range of native cell phenotypes and the deviation from it. As an example application to evaluating potential cell sources for cartilage or meniscus tissue engineering, we examine phenotypic changes of juvenile and adult articular chondrocytes and fibrochondrocytes across multiple passages and subsequent 3D culture. A substantial change was observed in cell phenotype due to the isolation process itself, followed by a clear progression toward the outer meniscal cell phenotype with passage. The new metrics also indicated that 3D culture moderately reduced the passage-induced deviation from the native meniscal phenotypes for juvenile chondrocytes and adult fibrochondrocytes, which was not obvious through examination of individual gene expressions. However, brief 3D culture alone did not move any of the cells towards an inner meniscal phenotype, the most relevant target for meniscal tissue engineering. This integrative approach of examining and combining multiple gene expressions can be used to evaluate various other tissue-engineering strategies to direct cells toward the desired phenotype. Copyright © 2015 John Wiley & Sons, Ltd.
authors.affiliation: Department of Bioengineering, Stanford University, CA, USA.
authors.forename: Min-Sun
authors.identifier: 0000-0002-4554-5659
authors.initials: MS
authors.lastname: Son
authors.affiliation: Department of Mechanical Engineering, Stanford University, CA, USA.
authors.forename: Marc E
authors.initials: ME
authors.lastname: Levenston
mesh_terms: D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D058948:Cell Tracking; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation
keywords: chondrocytes; fibrochondrocytes; gene expression; tissue engineering