Elucidating the contribution of the elemental composition of fetal calf serum to antigenic expression of primary human umbilical-vein endothelial cells in vitro

Abstract

One of the major obstacles to obtaining human cells of a defined and reproducible standard suitable for use as medical therapies is the necessity for FCS (fetal calf serum) media augmentation in routine cell culture applications. FCS has become the supplement of choice for cell culture research, as it contains an array of proteins, growth factors and essential ions necessary for cellular viability and proliferation in vitro. It is, however, a potential route for the introduction of zoonotic pathogens and makes defining the cell culture milieu impossible in terms of reproducibility, as the precise composition of each batch of serum not only changes but is in fact extremely variable. The present study determined the magnitude of donor variations in terms of elemental composition of FCS and the effect these variations had on the expression of a group of proteins associated with the antigenicity of primary human umbilical-vein endothelial cells, using a combination of ICPMS (inductively coupled plasma MS) and flow cytometry. Statistically significant differences were demonstrated for a set of trace elements in FCS, with correlations made to variations in antigenic expression during culture. The findings question in detail the suitability of FCS for the in vitro supplementation of cultures of primary human cells due to the lack of reproducibility and modulations in protein expression when cultured in conjunction with sera from xenogeneic donors.

Systematic review of factors influencing extracellular vesicle yield from cell cultures

“…Most in vitro culturing of cell monolayers requires the presence of serum in the culture medium to allow for the cells to adhere and grow, in which the most commonly used is FCS (Bryan et al 2011). The FCS has obvious inherent effects on the cultured cells, which varies between cell types and their differentiation status (Lund et al 2009;Bryan et al 2011).…”

“…The FCS has obvious inherent effects on the cultured cells, which varies between cell types and their differentiation status (Lund et al 2009;Bryan et al 2011). Since FCS contains vast amounts of EVs, most studies perform an EV depletion protocol of FCS before using it for production of CM, or change to serum-free medium (Shelke et al 2014).…”

Effect of fetal bovine serum on mineralization in silk fibroin scaffolds

“…It has been previously shown that differences in serum composition lead to statistical differences in experimental outcomes [40]. The disadvantages of FBS, such as undefined composition or batch-to-batch variations, have been known for decades.…”

“…Differences in spontaneous mineralization between the four FBS types used can be explained by differences in elemental composition. It is known that sera exhibit significant differences in composition between different types and batches [2,40]. The elemental composition of FBS, or sera in general, is not defined.…”

The Influence of Micronutrients in Cell Culture: A Reflection on Viability and Genomic Stability

“…The composition of these media includes certain vitamins and minerals, but unfortunately, in many common culture media, the only source of micronutrients is fetal bovine serum (FBS), which contributes to only 5–10% of the media composition. Moreover, the appropriate proportion of micronutrients is not always provided because the precise composition of each batch of FBS is in fact extremely variable [34]. …”