Cell expansion of human articular chondrocytes on macroporous gelatine scaffolds—impact of microcarrier selection on cell proliferation

Abstract

This study investigates human chondrocyte expansion on four macroporous gelatine microcarriers (CultiSpher) differing with respect to two manufacturing processes—the amount of emulsifier used during initial preparation and the gelatine cross-linking medium. Monolayer-expanded articular chondrocytes from three donors were seeded onto the microcarriers and cultured in spinner flask systems for a total of 15 days. Samples were extracted every other day to monitor cell viability and establish cell counts, which were analysed using analysis of variance and piecewise linear regression. Chondrocyte densities increased according to a linear pattern for all microcarriers, indicating an ongoing, though limited, cell proliferation. A strong chondrocyte donor effect was seen during the initial expansion phase. The final cell yield differed significantly between the microcarriers and our results indicate that manufacturing differences affected chondrocyte densities at this point. Remaining cells stained positive for chondrogenic markers SOX-9 and S-100 but extracellular matrix formation was modest to undetectable. In conclusion, the four gelatine microcarriers supported chondrocyte adhesion and proliferation over a two week period. The best yield was observed for microcarriers produced with low emulsifier content and cross-linked in water and acetone. These results add to the identification of optimal biomaterial parameters for specific cellular processes and populations.

PCL microspheres tailored with carboxylated poly(glycidyl methacrylate)–REDV conjugates as conducive microcarriers for endothelial cell expansion

“…24,25 Microcarriers have also been used 75 successfully as cell delivery systems to regenerate lost tissues, either applied directly via injection. 26 The various materials that microcarriers have been manufactured from include cellulose, gelatin, dextran, polyethylene and polystyrene. 26 The various materials that microcarriers have been manufactured from include cellulose, gelatin, dextran, polyethylene and polystyrene.…”

Past, present, and future of microcarrier-based tissue engineering

“…At the initial stage of microcarrier development, microcarriers could amplify cells and effectively maintain cell-specific phenotypes [43] . With gradual improvement in microcarrier culture technology and the application of biodegradable materials, microcarriers with tissue-engineered scaffolds have helped in the repair of damaged or degenerated tissue and the restoration of the original tissue structure and function.…”

Parallel fabrication of macroporous scaffolds

“…Macroporous scaffolds are three‐dimensional (3D) materials which contain inter‐connected networks of porous structures. Increased surface area‐per‐unit‐volume in these scaffolds facilitate higher numbers of binding sites for cellular and biomolecular adhesion compared to non‐porous scaffolds (Pettersson, Wetterö, Tengvall, & Kratz, ). In addition, the porous network facilitates transport of cells (Hollister, ), nutrients and metabolic wastes (Lee, Ahn, Chun, & Kim, ; Tan, Li, Ren, & Chen, ).…”