A Novel Flow‐Perfusion Bioreactor Supports 3D Dynamic Cell Culture

Abstract

Background. Bone engineering requires thicker three-dimensional constructs than the maximum thickness supported by standard cell-culture techniques (2 mm). A flow-perfusion bioreactor was developed to provide chemotransportation to thick (6 mm) scaffolds. Methods. Polyurethane scaffolds, seeded with murine preosteoblasts, were loaded into a novel bioreactor. Control scaffolds remained in static culture. Samples were harvested at days 2, 4, 6, and 8 and analyzed for cellular distribution, viability, metabolic activity, and density at the periphery and core. Results. By day 8, static scaffolds had a periphery cell density of 67% ± 5.0%, while in the core it was 0.3% ± 0.3%. Flow-perfused scaffolds demonstrated peripheral cell density of 94% ± 8.3% and core density of 76% ± 3.1% at day 8. Conclusions. Flow perfusion provides chemotransportation to thick scaffolds. This system may permit high throughput study of 3D tissues in vitro and enable prefabrication of biological constructs large enough to solve clinical problems.

Design and Functional Testing of a Multichamber Perfusion Platform for Three‐Dimensional Scaffolds

“…The 3-thread bayonet coupling, which connects the reservoir to the housing, makes the sterile assembling of the device fast and intuitive for the operator (a 1/4-turn bayonet is more immediate in laboratory use than O-ring couplings or screws-and-nuts fixations [10, 14, 18, 23]) and ensures a homogeneous compression of the deformable scaffold holder cartridge avoiding leakages. The reservoir, after mounting, is fluidically integrated within the culture chamber to reduce the dead priming volume of the overall circuit down to 3.5 mL, quite smaller than the 80 mL reported by Sailon et al [12] and comparable to the 3 mL stated by Domansky et al [26]. Priming volume may be increased up to 7 mL by adding medium in the reservoir.…”

Towards personalized medicine with a three-dimensional micro-scale perfusion-based two-chamber tissue model system

“…Cells cultured in such a 3D environment are likely to form complex structures that preserve cellular functions important in vivo . It is well known that cells in 3D respond better to microenvironmental cues and chemical or mechanical signals than those in 2D cell cultures for cell differentiation, proliferation, metabolism, and metastasis [46,51,59,60]. Also, cellular response to chemotherapeutic agents can be drastically different between 2D and 3D cultures, varying as much as 1000 folds [61].…”

The role of perfusion bioreactors in bone tissue engineering

“…This also limits the dimensions of the scaffolds that can be used on static culture 19 . In traditional two-dimensional culture techniques diffusion is enough to provide nutrients and oxygen to all the cells and waste removal, but when using three-dimensional constructs, diffusion is not sufficient 20 . Hence more complex bioreactor systems can be used to improve culture media circulation and convective transport of nutrients to the cells, allowing the development of a more uniform tissue.…”