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.

Combining TGF-β1 and Mechanical Anchoring to Enhance Collagen Fiber Formation and Alignment in Tissue-Engineered Menisci

“…In addition, in this study, fibrochondrocytes from juvenile bovine menisci were employed. It has been demonstrated that fibrochondrocytes from adult bovids show a higher ratio of collagen type I to II gene expression compared to those cells from juvenile bovids . The effects of growth factors on fibrochondrocytes from either juvenile or adult bovids also vary .…”

“…It has been demonstrated that fibrochondrocytes from adult bovids show a higher ratio of collagen type I to II gene expression compared to those cells from juvenile bovids. 82 The effects of growth factors on fibrochondrocytes from either juvenile or adult bovids also vary. 19 With regard to the zonal difference, it has been shown that cells from the inner zone produce more proteoglycans than those cells from the outer zone.…”

Surgical and tissue engineering strategies for articular cartilage and meniscus repair

“…Although meniscal fibrochondrocytes might seem to be an obvious choice for engineering the meniscus, co-culturing these cells with others might be required to achieve the best results. Similar to chondrocytes, meniscal fibrochondrocytes dedifferentiate when expanded 135 , a fact that has led to the investigation of MSCs from the bone marrow 136 , synovium 137 and adipose tissue as alternative cell sources 138 . In a 2017 study,…”

Advances in Regenerative Sports Medicine Research

“…While scaffolds are beneficial due to their ability to incorporate growth factors and their initial mechanical stability, they also indicate recapitulation of the mechanical and biochemical architecture of the native meniscus, with matched stiffness and ingredient gradients (Higashioka, et al, 2014;Steele, et al, 2014;Zhu, et al, 2018;Zitnay, et al, 2018). Notably, when engineering menisci for regeneration, although an obvious choice for cell source might be fibrochondrocytes, regeneration effects are best when fibrochondrocytes are cocultured with other cell subsets (Hadidi, et al, 2016;Koh, et al, 2017;Son and Levenston, 2017;Sasaki, et al, 2018;Xie, et al, 2018). Zhang C. H. et al (2018) used a pre-mechanically stimulated poly (ε-caprolactone) (PCL) scaffold, cocultured with rabbit bone marrow stem cells, for meniscus replacement and found that the pretreated scaffold was a better choice for inducing tissue regeneration (Figure 2).…”