Three-dimensional cell culture and tissue engineering in a T-CUP (Tissue Culture Under Perfusion)

The aim of this study was to develop and validate a simple and compact bioreactor system for perfusion cell seeding and culture through 3- dimensional porous scaffolds. The developed Tissue Culture Under Perfusion ( T- CUP) bioreactor is based on the concept of controlled and confined alternating motion of scaffolds through a cell suspension or culture medium, as opposed to pumping of the fluid through the scaffolds. Via the T- CUP, articular chondrocytes and bone marrow stromal cells could be seeded into porous scaffolds of different compositions and architectures ( chronOS(TM), Hyaff (R)- 11, and Polyactive(TM)) at high efficiency ( greater than 75%), uniformity ( cells were well distributed throughout the scaffold pores), and viability ( greater than 97%). Culture of articular chondrocytes seeded into 4- mm thick Polyactive(TM) scaffolds for 2 weeks in the T- CUP resulted in uniform deposition of cartilaginous matrix. Cultivation of freshly isolated human bone marrow nucleated cells seeded into ENGipore ceramic scaffolds for 19 days in the T- CUP resulted in stromal cell- populated constructs capable of inducing ectopic bone formation in nude mice. The T- CUP bioreactor represents an innovative approach to simple, efficient, and reliable 3D cell culture, and could be used either as a model to investigate mechanisms of tissue development or as a graft manufacturing system in the context of regenerative medicine.