Development of a serum free medium for human diploid fibroblasts

  • Raes, Martine (PI)
  • Eliaers, Francois (Technician)
  • VERTEZ, Marie-Jeanne (Technician)

Project: Research

Project Details

Description

The aim of our work is to formulate a serum free medium for MRC-5 cells (human diploid fibroblasts from fetal lung) which are commonly used in the industrial field for the manufacture of biologicals.
Animal serum (mainly bovine) is essential for cell culture because it is an important source of nutrients, growth and attachment factors, protection agents' It is also used to inhibit trypsin in subculture protocol and for cryopreservation. However, it presents several disadvantages especialy for the pharmaceutical industry :
- substantial cost,
- variability between batches,
- serum is an undefined mixture,
- poor reliability of serum supplying,
- last but not least, it is a source of pathogenic contaminants (adventitious agents) such as fungi, bacteria, viruses or agents of the bovine spongiform encephalopathy (BSE - until now, no detectable infectivity was found in serum, but it remains a theoritical risk).
Therefore, the current tendency is to recommend the elimination of serum and even more the elimination of all the animal-derived raw material from the culture media used in the manufacture of biologicals for human therapy or vaccination.
Several serum free media have already been developed for transformed cell lines (e.g. CHO, hybridoma or recombinant myeloma cells) but nothing is available for normal cells. Our project focuses on human diploid MRC-5 cells. Because of their limited life-span, these cells have particular requests on the quality of the culture medium.
We are developing a long term culture protocol (several weeks) in which serum is replaced at the level of the culture medium, trypsin inhibition and the cryopreservation medium. After a preliminar screening to identify the essential factors for the MRC-5 cell proliferation, a prototype medium is now tested in long term cultures and the cell culture conditions are under optimalization. Afterwards, the cells adapted to the serum free medium will be further characterized at the molecular level.
StatusFinished
Effective start/end date1/04/9831/03/01

Keywords

  • cell culture
  • diploid cells
  • fibroblasts
  • serum free medium