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Towards a new in vitro model for research in Tissue Mechanobiology
The use of 3D culture models has increased considerably in the past years. For reasearch in tissue engineering as well as in mechanobiology, stable culture conditions are crucial.
Keywords: explant culture, 3d culture, tissue culture, musculoskeletal soft tissue, tissue engineering, in vitro, biology
In order to develop a functional engineered soft tissue implant, an in depth understanding of the structure and function of the natural tissue is required. As an in vitro model to study cell response to mechanical loading, an explant culture system is often used (the culture of tissue fragments in growth medium). Explant culture has the advantage over other culture techniques that cells are left in their natural surrounding extracellular matrix, which more accurately mimics the in vivo environment.
Preliminary data already showed that our tissue explants can be cultured in standard culture medium conditions for at least 3 weeks, with high cell viability and structural integrity maintained. However, with this project we want to go one step further, and identify conditions under which culture in chemically-defined medium without serum supplementation is possible. The supplementation of culture medium with serum (FBS/FCS, “fetal bovine serum”/“fetal calf serum”) is still common practice in cell culture applications. It contains essential constituents for cell proliferation and maintenance such as hormones, vitamins, transport proteins, trace elements, spreading and growth factors. However, due to a number of disadvantages in terms of quality and reproducibility of in vitro data, heterogeneous influence of FBS/FCS on cells and mechanical tissue properties, animal welfare concerns, and constantly growing market demand, the search for alternatives and the development of serum-free medium formulations has gained attention.
In order to develop a functional engineered soft tissue implant, an in depth understanding of the structure and function of the natural tissue is required. As an in vitro model to study cell response to mechanical loading, an explant culture system is often used (the culture of tissue fragments in growth medium). Explant culture has the advantage over other culture techniques that cells are left in their natural surrounding extracellular matrix, which more accurately mimics the in vivo environment. Preliminary data already showed that our tissue explants can be cultured in standard culture medium conditions for at least 3 weeks, with high cell viability and structural integrity maintained. However, with this project we want to go one step further, and identify conditions under which culture in chemically-defined medium without serum supplementation is possible. The supplementation of culture medium with serum (FBS/FCS, “fetal bovine serum”/“fetal calf serum”) is still common practice in cell culture applications. It contains essential constituents for cell proliferation and maintenance such as hormones, vitamins, transport proteins, trace elements, spreading and growth factors. However, due to a number of disadvantages in terms of quality and reproducibility of in vitro data, heterogeneous influence of FBS/FCS on cells and mechanical tissue properties, animal welfare concerns, and constantly growing market demand, the search for alternatives and the development of serum-free medium formulations has gained attention.
The goal of this project is to investigate various conditions for serum free culture and to show that the conditions are suitable for culture over several weeks, e.g. by describing proliferation and cell viability using different assays such as live/dead fluorescent staining and confocal microscopy, and maintenance of matrix composition and integrity using histology.
The goal of this project is to investigate various conditions for serum free culture and to show that the conditions are suitable for culture over several weeks, e.g. by describing proliferation and cell viability using different assays such as live/dead fluorescent staining and confocal microscopy, and maintenance of matrix composition and integrity using histology.
If you are interested, please send an e-mail to stephanie.huber@hest.ethz.ch
More information about our lab: http://www.orthotech.ethz.ch/
If you are interested, please send an e-mail to stephanie.huber@hest.ethz.ch
More information about our lab: http://www.orthotech.ethz.ch/