Adaptation of the HEp-2 cell line to totally animal-free culture systems and real-time analysis of cell growth

Abstract

Routine cell culture demands the use of animal-derived products, mainly fetal bovine serum and swine or bovine trypsin. According to the 3Rs principle and to the European Centre for the Validation of Alternative Methods, animal-free substitutes are strongly recommended for in vitro methods. In this study, the HEp-2 cell line was adapted to different totally animal-free culture systems, such as a serum-free complete medium (VP-SFM), human platelet lysate and a synthetic trypsin (TrypLE™ Express); afterward, cell growth was assessed with the xCELLigence instrument. Animal-free products provided promising results, with performances similar or preferable to the common reagents; therefore their use could be encouraged for both ethical and technical advantages.

METHOD SUMMARY

HEp-2 cells were sequentially adapted to the serum-free complete medium VP-SFM, performing subcultures into 25, 50, 75, 90, 95, 98, 99 and 100% VP-SFM. A direct adaptation to the medium was achieved with 5% platelet lysate. TrypLE™ Express was employed as a direct replacement of animal trypsin for the detachment of HEp-2 cells, without the use of an inhibitor. Real-time analysis of cell growth and proliferation and the comparison of different culture conditions were determined with the xCELLigence method.