An Effect of Culture Media on Epithelial Differentiation Markers in Breast Cancer Cell Lines MCF7, MDA-MB-436 and SkBr3

Abstract

Background and objectives: Cell culture is one of the mainstays in the research of breast cancer biology, although the extent to which this approach allows to preserve the original characteristics of originating tumor and implications of cell culture findings to real life situations have been widely debated in the literature. The aim of this study was to determine the role of three cell culture media on transcriptional expression of breast cancer markers in three breast cancer reference cell lines (MCF7, SkBr3 and MDA-MB-436). Materials and methods: Cell lines were conditioned in three studied media (all containing 5% fetal bovine serum (FBS) + hormones/growth factors; different composition of basal media) for four passages. Population growth was characterized by cumulative population doubling levels, average generation time, cell yield and viability at the fourth passage. Transcriptional expression of breast cancer differentiation markers and regulatory transcriptional programs was measured by qPCR. Results: Differences in the composition of growth media significantly influenced the growth of studied cell lines and the expression of mammary lineage governing transcriptional programs and luminal/basal markers. Effects of media on transcriptional expression were more pronounced in luminal cell lines (MCF7, SkBr3), than in the basal cell line (MDA-MB-436). Changes in growth media in terms of supplementation and basal medium delayed growth of cells, but improved cell yields. Conclusions: The expression of breast cancer cell differentiation phenotypic markers depends on the composition of cell growth medium, therefore cell culture as a tool in phenotypic studies should be used considering this effect. The findings of such studies should always be interpreted with caution. The formulation of cell growth media has greater effect on the expression of phenotypic markers in luminal, rather than basal cell lines. Media containing mitogens and higher vitamin content improved efficacy of cell culture in terms of cell yields, although greatly increased growth times.

Selecting suitable reference genes for qPCR normalization: a comprehensive analysis in MCF-7 breast cancer cell line

“…Culture and seeding conditions of MCF-7 biological replicate cultures (sub-clones A1 and A2) Samples were collected from MCF-7 cell line (ATCC, HTB-22) that has been used in our laboratory for previous studies [56]. Samples from different passages were cryopreserved during culturing at different time points.…”

“…The MCF-7 samples from previous study in our laboratory [56] were used to validate the reference gene triplet in cells undergoing different nutrient conditions, i.e. cultured in different media.…”

A novel 20-gene prognostic score in pancreatic adenocarcinoma

“…Keratinocyte differentiation, skin development and epidermis development gene sets enriched in the high PPS20 group include many genes belonging to the keratin family, among which KRT16 has been used as a basal cell marker [28,29]; KRT17 has been shown to induce cancer stem cell-like properties in cervical cancer [30] and tumor growth, motility and invasion in gastric cancer [31], which is also associated with poor prognosis in breast cancer [32]. Formation of primary germ layer and endoderm gene sets were also enriched in tumors with high PPS20.…”

Co-occurrence of Cell Lines, Basal Media and Supplementation in the Biomedical Research Literature

“…Several studies have shown that the composition of basal medias has a significant impact on the results of their experiments (Ariffin et al, 2016;Kim et al, 2015;Pirsko et al, 2018;Selenius et al, 2019;Tomoya Kawakami, 2016). However, culture media and supplementation remain a largely overlooked experimental parameter.…”