Moreira Teixeira L.S. - High Throughput Generated Micro-Aggregates of Chondrocytes Stimulate Cartilage Formation In Vitro and In Vivo

L.S. Moreira Teixeira.1,§, J.C.H. Leijten1,§, J. Sobral2, R. Jin3, A.A. van Apeldoorn1, J. Feijen3, C. van Blitterswijk2, P.J. Dijkstra1,3 and M Karperien1,*

European Cells and Materials 2012

Abstract

Cell-based cartilage repair strategies such as matrixinduced autologous chondrocyte implantation (MACI) could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous matrix deposition and remodelling. To address this issue, we designed a micromould to enable controlled high-throughput formation of micro-aggregates. Morphology, stability, gene expression profiles and chondrogenic potential of micro-aggregates of human and bovine chondrocytes were evaluated and compared to single-cells cultured in micro-wells and in 3D after encapsulation in Dextran-Tyramine (Dex-TA) hydrogels in vitro and in vivo. We successfully formed micro-aggregates of human and bovine chondrocytes with highly controlled size, stability and viability within 24 hours. Micro-aggregates of 100 cells presented a superior balance in Collagen type I and Collagen type II gene expression over single cells and micro-aggregates of 50 and 200 cells. Matrix metalloproteinases 1, 9 and 13 mRNA levels were decreased in micro-aggregates compared to singlecells. Histological and biochemical analysis demonstrated enhanced matrix deposition in constructs seeded with micro-aggregates cultured in vitro and in vivo, compared to single-cell seeded constructs. Whole genome microarray analysis and single gene expression profiles using human chondrocytes confirmed increased expression of cartilagerelated genes when chondrocytes were cultured in microaggregates. In conclusion, we succeeded in controlled highthroughput formation of micro-aggregates of chondrocytes. READ MORE.....