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.
Highlights
Long-lived cancer cell lines, which have been derived from patient tumor cells, have been widely used to explore biology of breast cancer and new therapies in the past and currently remain one of the main models for investigation of breast cancer [1]
Most molecular profiling studies of breast cancer cell lines have been conducted after culture of cells in medium traditionally used for the breast cancer cell line in the particular laboratory, e.g., in the study of expression profiling of large panel of breast cancer cell lines and in vivo breast tumors by Prat et al [6]
MCF7 and SkBr3 cell lines were cultured in RPMI-1640 with 10% fetal bovine serum (FBS) and the MDA-MB-436 cell line was cultured in DMEM with 10% FBS, while in the study of genomic and transcriptional characterization of breast cancer cell lines by Neve et al [7], the same cell lines were grown in DMEM medium, McCoy 5A medium and L15 medium, respectively—all media with 10% FBS
Summary
Long-lived cancer cell lines, which have been derived from patient tumor cells, have been widely used to explore biology of breast cancer and new therapies in the past and currently remain one of the main models for investigation of breast cancer [1]. 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. Conclusions: The expression of breast cancer cell differentiation phenotypic markers depends on the composition of cell growth medium, 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. Media containing mitogens and higher vitamin content improved efficacy of cell culture in terms of cell yields, greatly increased growth times
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