Abstract
The use of fetal bovine serum hinders obtaining reproducible experimental results and should also be removed in hormone and growth factor studies. In particular hormones found in FBS act globally on cancer cell physiology and influence transcriptome and metabolome. The aim of our study was to develop a renal carcinoma serum free culture model optimized for (embryonal) renal cells in order to select the best study model for downstream auto-, para- or endocrine research. Secondary aim was to verify renal carcinoma stem cell culture for this application. In the study, we have cultured renal cell carcinoma primary tumour cell line (786-0) as well as human kidney cancer stem cells in standard 2D monolayer cultures in Roswell Park Memorial Institute Medium or Dulbecco’s Modified Eagle’s Medium and Complete Human Kidney Cancer Stem Cell Medium, respectively. Serum-free, animal-component free Human Embryonic Kidney 293 media were tested. Our results revealed that xeno-free embryonal renal cells optimized culture media provide a useful tool in RCC cancer biology research and at the same time enable effective growth of RCC. We propose bio-mimic RCC cell culture model with specific serum-free and xeno-free medium that promote RCC cell viability.
Highlights
Fetal bovine serum (FBS) is a component of high activity added to basic medium including RPMI (Roswell Park Memorial Institute medium), DMEM (Dulbecco’s Modified Eagle’s Medium), MEM (Minimum Essential Medium) or McCoy0s 5A or Ham’s F-12 media in order to promote cell growth and proliferation
In standard 2D cell culture in regular T-75 flasks we have observed that HKCSC cells proliferated faster in normoxia (Fig. 1a, b, e), while 786-0 cells showed higher proliferation rate in hypoxia over first 3 days and proliferation rate decreased (Fig. 1c, d, f, g), as defined both by MTT and Alamar Blue reduction
FBS can support the growth of many cell types including Renal cell cancer (RCC), how it does so and which specific factors are important for each individual cell type has not been identified until now
Summary
FBS is a mixture of multiple compounds and proteins It is characterized by a high content of embryonic growth promoting factors along with many defined and undefined components— macromolecular proteins, low molecular weight nutrients, carrier proteins for water-insoluble components, hormones, attachment factors, spreading factors and low amount of gamma globulins (Shah 1999; Paschoal et al 2014). It is widely accepted that composition of FBS directly influences the outcome of cellular experiments. It introduces the greatest variability into cell culture in experiment to experiment and between laboratories. Using FBS to culture specific cells in vitro is non-physiological and may lead to aberrant results in any experiment because organ-specificity is not kept (Maggs et al 1995; Bhave and Neilson 2011)
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