#1949 Comparison of two commercial available primary human mesangial cell clones reveal differences

Abstract

Abstract Background and Aims Mesangial cells and their matrix form the central stalk of the glomerulus but is often overlooked when studying glomerular function. The cells are however highly involved in glomerular function and crosstalk between the different cell types of the glomerulus. They are important in onset and progression of several glomerular diseases, such as IgA nephropathy and diabetic kidney disease. As mesangial cells have several functions and are involved in glomerular disease they are often used for in vitro work studying response to various stimuli and downstream signaling. This includes changes in the production and regulation or their matrix, proliferation, contractility, protein expression and release of different inflammatory mediators such as cytokines and growth factors [1]. Using primary cells when doing in vitro work is usually considered the best option, but there might be differences between origin of cells in their response and physiology. For this study, our aim was to compare two types of commercially available primary human mesangial cells from Cell systems (CS) and Novabiosis (NBS). Method For characterization of the CS and NBS mesangial cells, gene expression of commonly used mesangial cell markers, including KRT19, ACTA2, PDGFRA and PDGFRB, was analyzed with real time qPCR. Staining of smooth muscle actin was done in order to compare the phenotype of CS and NBS. To compare the cells, differences in protein expression were determined using proteomics. CS and NBS Cells were cultured in the same way and were also treated with PDGF-BB for 24 h to compare with untreated control. For further comparison, proliferation assay was used to compare proliferation after treating cells with PDGF-BB for 24 h. Results Both sources of cells expressed all markers, but there was a difference in level of gene expression. CS cells showed higher expression than NBS cells for all markers, apart from ACTA2. NBS cells had a more stellate phenotype whereas the CS cells were more elongated. There was a difference in proteins expressed by CS and NBS mesangial cells from our proteomics. The level of upregulation of proteins seen with proteomics was stronger for CS than NBS cells when treated with PDGF-BB. The number of proteins upregulated was also higher for CS than NBS cells. Both mesangial cell types had increased proliferation when treated with PDGF-BB, however CS cells were seen to grow faster in cell culture and seemed to be more responsive to the treatment with PDGF-BB. Conclusion In conclusion, this study compares two commercially available primary human mesangial cells and show that there is a difference between the two. The cell types react differently when treated with PDGF-BB and has some difference in what proteins they express. This needs to be considered when conducting cell culture experiments with mesangial cells, depending on what the aim and focus for the study is.