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, Hyaff-11, and Polyactive) 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 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.
Timmins
Nicholas E
NE
Tissue Engineering, Laboratory 405, Departments of Surgery and of Research, Hebelstrasse 20, CH-4031 Basel, Switzerland.
Scherberich
Arnaud
A
Früh
Jennifer-Annika
JA
Heberer
Michael
M
Martin
Ivan
I
Jakob
Marcel
M
D000818:Animals; D019149:Bioreactors; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation; D002358:Cartilage, Articular / Q000166:cytology; D002417:Cattle; D018929:Cell Culture Techniques / Q000295:instrumentation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D010477:Perfusion; D046509:Tissue Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods*