Skip to main content
European Commission logo print header

Development of an immortalised human articular cartilage cell line and its use in physiological, pharmacological and toxicological in vitro investigation

Objective

The objectives are the development of an immortalized human articular cell line and investigation of its suitability in physiological, pharmacological and toxicological in vitro studies.
The research project was to study the possible use of invitro cultured phenotypically stable human cartilage cells (chondrocytes) in pathophysiological, pharmacological and toxicological investigations. Such assay systems would be invaluable in the study of human diseases such as the destruction of joints in rheumatic disorders and beneficial and deleterious effects of pharmacological substances on cartilage cells. The need of using experimental animal models could be partly obviated.

Cartilage cells were obtained from human subjects with spontaneously occurring degenerative and inflammatory joint diseases. Since such cells are difficult to obtain, it was decided to construct immortalized human chondrocyte cell lines. Because of a variety of control mechanisms the genetic apparatus of the human cell is relatively stable compared to that of most other animal species. It was necessary to study the problems inherent in the immortalization and propagation of the human chondrocyte in particular and of human cells in general. To this end, several clones of immortalized human cartilage cells had to be developed.

Several specimens of human articular chondrocytes were isolated at autopsy from articular cartilage of human femoral condyles. The investigation resulted in: development of a protocol for biological freezing and preservation of viable chondrocytes at -180 C;
characterization of the metabolic functions of the human chondrocyte cultured in suspension culture in agarose;
standardization of the culture conditions;
study of the influences of various biological mediators and some pharmacological agents on chondrocytes;
definition of the nonimmortalized cartilage cell;
studies of the suitability of this cell for research on cell cell (macrophage connective tissue cell) interactions in pathological conditions;
development of immortialized chondrocyte clones orginating from different human subjects.

To the best of knowledge these are the first human articular cartilage cell (chondrocyte) lines. The cells are cultured as phenotypically stable articular cartilage chondrocytes in a suspension culture system. They export extracellular matrix products typical of articular cartilage. A major difficulty encountered was that after the initial transformation the cells remained in a 6 to 9 month long latency or crisis period.
The aim of the study is the development of an immortalized human cartilage cell line, and the investigation of its function with respect to the homeostasis of extracellular matrix macromolecules. The preservation of the original (in vivo) functions in an in vitro model (culture in different gelified matrices) will be investigated. Functional characterisation of this cell line will involve the study of synthesis, secretion and turnover of proteoglycan and collagen in an artificial extracellular matrix. In order to obtain full control of the environment and complete standardisation of this system, insulin-like growth factor will replace serum in the incubation medium.

The system will be used to explore the ways in which normal and altered chondrocytes build, degrade and/or repair their environment. The effects of disease mediators (cytokines, oxide radicals, co-cultured macrophages) in this model, as well as the effects of some 'pilot' drugs (steroids, non-steroidal anti-inflammatory drugs, interleukin-1-inhibitors) on mediated chondrocyte metabolism will be studied. To this end cells will be obtained from human donors (autopsy). Finally, cartilage cells will be purchased from experimental animal models of degenerative and inflammatory joint diseases in order to determine whether in vitro observed abnormalities of these cells are consistent with the pathological changes encountered in vivo, and whether their phenotypes are preserved after immortalisation.

Topic(s)

Data not available

Call for proposal

Data not available

Coordinator

Universiteit Gent
EU contribution
No data
Address
185,De Pintelaan
9000 Gent
Belgium

See on map

Total cost
No data

Participants (4)