Development of a serum free medium for a human diploid fibroblast cell line

摘要

The aim of this work was to formulate a serum-free medium (or a medium containing a reduced serum concentration) for MRC-5 cells (human diploid fibroblasts from foetal lung. The screening was performed in multi-well plates using the propidium iodide assay to follow cell proliferation. Among all the media tested, the prototype medium B (containing 0.5% foetal bovine serum (FBS) and several growth factors and hormones) stimulated the proliferation of MRC-5 cells in a similar way to our reference medium (Ultra-MEM + 2% FBS). The quality of medium B was confirmed by the incorporation of 5-bromo-2'-deoxyufidine (BrdU) in newly synthesised DNA. This medium also supported MRC-5 cell growth over 8 population doublings. Finally, medium B can be slightly improved by the addition of the synthetic peptide cyclo(GRGDSPA) acting through the interaction with integrins, confirming the hypothesis of a crosstalk between the integfin and the growth factor receptor activation pathways.