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Alternatives to Fetal Calf Serum: Thrombocyte extracts for the substitution of fetal serum in cell- and tissue culture

Project Aim

The aim of the proposed research project is to investigate the growth and proliferation effects of human thrombocyte extracts on cells cultivated in vitro, and to develop thrombocyte extracts as a true alternative to fetal calf serum in cell and tissue culture.

This project has high biotechnological potential for commercial use in production and distribution of human thrombocyte extracts for the replacement of fetal serum in cell and tissue culture as well as in tissue engineering.

Project Background

Addition of serum to the cell culture medium is essential for the optimal in vitro cultivation of eucaryotic cells, i.e. growth, proliferation and cell specific differentiation. Serum supplies hormones and growth factors, binding proteins and anchoring factors, as well as vitamins and trace elements. In addition to animal sera of different developmental stages (fetal, new born and adult), sera of diverse species are also used. The use of serum also has a number of disadvantages: (1) Serum is an undefined supplement, the exact qualitative composition is not yet known; the quantitative composition also varies. (2) Severe doubts recently arose concerning the possible suffering the calves may endure during raw serum extraction (van der Valk et. Al., 2004). (3) Not long ago, questions of concern were even raised about whether the world-wide demand for fetal calf serum for research and biotech-industry can actually be met. To avoid these disadvantages and to generate defined and controlled culture conditions, extensive efforts are being undertaken to find adequate alternatives to the use of this serum in cell culture (Falkner et al., 2006; Gstraunthaler, 2003).

As is generally known, growth stimulating and mitogenic effects can only be accomplished with the addition of serum, while the addition of plasma as a supplement is not adequate. This implies, and it was also concluded that the thrombocyte-factors PDGF, FGF, TGF-b and VEGF which are released during the blood coagulation, do have significant impact on the development of cells in vitro. It should furthermore be clarified to which extant thrombocyte extract is an adequate alternative to fetal calf serum in cell and tissue culture (Rauch et al., 2008).

Project duration

May 1st, 2006 - Decembre 31st, 2010

Participating Laboratories

The project is carried out at the Commission of Physiology at the Medical University of Innsbruck by Maga. Caroline Rauch und Drin. Elisabeth Feifel under the direction of Univ.-Prof. Dr. Gerhard Gstraunthaler

Financing

Pollux Foundation

References

Falkner E., Appl H., Eder C., Losert U.M., Schöffl H. and Pfaller W. Serum free cell culture: The free access online database. Toxicol. In Vitro 20: 395-400, 2006.

Gstraunthaler G. Alternatives to the use of fetal bovine serum: Serum-free cell culture. ALTEX 20: 275-281, 2003.

Rauch C., Feifel E., Schöffl H., Pfaller W., Gstraunthaler G. Alternatives to the use of fetal bovine serum: Platelet lysates as serum replacement in cell and tissue culture. ALTEX 24(4): 353, 2007.

van der Valk J., Mellor D., Brands R., Fischer R., Gruber F., Gstraunthaler G., Hellebrekers L., Hyllner J., Jonker F.H., Prieto P., Thalen M. and Baumans V. The humane collection of fetal bovine serum and possibilities for serum-free cell and tissue culture. Toxicol. In Vitro 18: 1-12, 2004.

van der Valk J., Brunner D., De Smet K., Fex Svenningsen Å., Honegger P., Knudsen L.E., Lindl T., Noraberg J., Price A., Scarino M.L. and Gstraunthaler G. Optimization of chemically defined cell culture media – Replacing fetal bovine serum in mammalian in vitro methods. Toxicol. in Vitro 24: 1053-1063, 2010.

Brunner D., Frank J., Appl H., Schöffl H., Pfaller W.and Gstraun-thaler G. Serum-free Cell Culture: The Serum-free Media interactive online database. ALTEX 27 (1): 53-62, 2010

Rauch C. Human Platelet Lysates as a Serum Substitute in Cell and Tissue Culture (PhD Thesis), Innsbruck Medical University, Dept. of Physiology, 2010