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Extracellular Matrix Stability of Primary Mammalian Chondrocytes and Intervertebral Disc Cells Cultured in Alginate-Based Microbead Hydrogels

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Abstract:

Three-dimensional alginate constructs are widely used as carrier systems for transplantable cells. In the present study, we evaluated the chondrogenic matrix stability of primary rat chondrocytes and intervertebral disc (IVD) cells cultured in three different alginate-based microbead matrices to determine the influence of microenvironment on the cellular and metabolic behaviors of chondrogenic cells confined in alginate microbeads. Cells entrapped in calcium, strontium, or barium ion gelled microbeads were monitored with the live/dead dual fluorescent cell viability assay kit and the 1,9-dimethylmethylene blue (DMB) assay designed to evaluate sulfated glycosaminoglycan (s-GAG) production. Expression of chondrogenic extracellular matrix (ECM) synthesis was further evaluated by semiquantitative RT-PCR of sox9, type II collagen, and aggrecan mRNAs. Results indicate that Ca and Sr alginate maintained significantly higher population of living cells compared to Ba alginate (p < 0.05). Production of s-GAG was similarly higher in Ca and Sr alginate microbead cultures compared to Ba alginate microbeads. Although there was no significant difference between strontium and calcium up to day 14 of culture, Sr alginate showed remarkably improved cellular and metabolic activities on long-term cultures, with chondrocytes expressing as much as 31% and 44% greater s-GAG compared to calcium and barium constructs, respectively, while IVD cells expressed 63% and 74% greater s-GAG compared to calcium and barium constructs, respectively, on day 28. These findings indicate that Sr alginate represent a significant improvement over Ca- and Ba alginate microbeads for the maintenance of chondrogenic phenotype of primary chondrocytes and IVD cells.
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  • Cell Transplantation publishes original, peer-reviewed research and review articles on the subject of cell transplantation and its application to human diseases. To ensure high-quality contributions from all areas of transplantation, separate section editors and editorial boards have been established. Articles deal with a wide range of topics including physiological, medical, preclinical, tissue engineering, and device-oriented aspects of transplantation of nervous system, endocrine, growth factor-secreting, bone marrow, epithelial, endothelial, and genetically engineered cells, among others. Basic clinical studies and immunological research papers are also featured. To provide complete coverage of this revolutionary field, Cell Transplantation will report on relevant technological advances, and ethical and regulatory considerations of cell transplants. Cell Transplantation is now an Open Access journal starting with volume 18 in 2009, and therefore there will be an inexpensive publication charge, which is dependent on the number of pages, in addition to the charge for color figures. This will allow work to be disseminated to a wider audience and also entitle the corresponding author to a free PDF, as well as prepublication of an unedited version of the manuscript.
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