Bioreactor cultivation of three-dimensional cartilage-carrier-constructs

Abstract

A flow-chamber bioreactor was designed for generation of three-dimensional cartilage-carrier-constructs. A specific attribute of the flow-chamber is a very thin medium layer for improved oxygen supply and a counter current flow of medium and gas. Three-dimensional cartilage-carrier-constructs were produced according to a standard protocol from chondrocytes of an adult mini-pig. The final step of this protocol was performed either in the bioreactor or in 12-well plates. The bioreactor experiments showed a significantly higher matrix thickness but a lower ratio of glycosaminoglycan to DNA. For both culture methods the constructs contained a high amount of collagen II. Appearance of the cartilage obtained in the bioreactor seemed to be closer to native cartilage with respect to distribution of the cells within the matrix, smoothness of the surface etc. All results considered the flow-chamber bioreactor is a very useful tool for generation of three dimensional cartilage-carrier constructs.

Electrical Stimulation of NIH-3T3 Cells with Platinum-PEDOT-Electrodes Integrated in a Bioreactor

“…In order to facilitate the exposure of cells to electric fields, the design of the perfusion bioreactor introduced by Nagel-Heyer et al . [12] was modified. The bioreactor for electrical stimulation consisted of a transparent base plate and a 24 well equivalent top frame as seen in Fig.…”

Redifferentiation of chondrocytes and cartilage formation under intermittent hydrostatic pressure

“…The cells were isolated from the knee-joint, which was kindly provided by a local slaughter. Cartilagecarrier constructs were generated according to a 4-step-protocoll suggested by Nagel-Heyer et al (2005). Differing from that protocol redifferentiation and cartilage formation was performed under an atmosphere of 5% (v/v) O 2 and 5% (v/v) CO 2 .…”

“…Collagen type I and II determination after cartilage formation was performed analogous to Nagel-Heyer et al (2005). Determination of collagen type II and I was performed separately via specific binding of monoclonal antibodies of mice against collagen type I (Ig G2a/k) and collagen type II (IgG1/k) and binding of the biotinylated secondary antibody (IgG (H + L)-biotin) Goat anti-mouse (Southern Biotech).…”

A Differential Pressure Laminar Flow Reactor Supports Osteogenic Differentiation and Extracellular Matrix Formation from Adipose Mesenchymal Stem Cells in a Macroporous Ceramic Scaffold

“…20 A fixed-bed perfusion chamber, developed by the group of Pörtner for cartilage engineering, allowed transverse perfusion from above the cylindrical scaffolds, situated in small cavities, with a counter-current flow of gas on top of the medium. 21 Another interesting approach has been described by Li et al, where a customized porous scaffold, with an inner blind-ending tunnel, was perfused through the central tunnel toward the pores while being surrounded by a culture medium in a reservoir. 22 A perfusion system by Zhao et al, modified by Kim and Ma, allows parallel (horizontal) or transverse flow through three scaffolds placed in a row.…”