ALTEX :: Alternatives to Animal Experiments

2004, VOLUME 2

Culturing Cells Without Serum: Lessons Learnt Using Molecules of Plant Origin

Patricia Pazos, Monica Boveri, Alessandra Gennari, Juan Casado, Fernando Fernandez and Pilar Prieto

ECVAM, Institute for Health and Consumer Protection, Joint Research Centre, European Commission, I-Ispra

SUMMARY

To successfully grow cells in serum-free medium is an interesting challenge to cell biology. The use of such media for in vitro cell culture work would be a key contribution to the 3Rs concept, enabling the avoidance of the use of animals and animal products at all stages of the experiment. In addition, numerous problems related to virus infections transmitted by animal serum would be avoided, thus increasing reproducibility. Prolifix is a new reagent of plant origin. It contains a molecule (GCR 1003) that has an activity similar to that of the mitogenic molecules in serum and could be suitable to substitute serum in culture medium. Two epithelial cell lines, LLC-PK1 and Caco-2, were progressively adapted to a special culture medium containing 10% Prolifix in the absence of serum. After adaptation, cell cultures were characterised. We found that these reagents of plant origin could be promising alternatives to serum. However, more work and effort is needed to improve cell adaptation, cell attachment, growth rates as well as freezing/thawing protocols.

Keywords: serum, replacement, in vitro, epithelial cells, plant components