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

机译:培养基对乳腺癌细胞MCF7MDA-MB-436和SkBr3上皮分化标记的影响

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摘要

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.
机译:背景与目的:细胞培养是乳腺癌生物学研究的主要内容之一,尽管这种方法在多大程度上可以保留起源肿瘤的原始特征以及细胞培养结果对现实生活状况的影响,在2004年已被广泛争论。文献。这项研究的目的是确定三种细胞培养基对三种乳腺癌参考细胞系(MCF7,SkBr3和MDA-MB-436)中乳腺癌标志物转录表达的作用。材料和方法:在三种研究培养基(均含有5%胎牛血清(FBS)+激素/生长因子;基础培养基的组成不同)中调节细胞系4代。种群增长的特征是累积的种群倍增水平,平均世代时间,细胞产量和第四代的活力。通过qPCR测量乳腺癌分化标志物和调控转录程序的转录表达。结果:生长培养基组成的差异显着影响了所研究细胞系的生长以及控制转录程序和腔/基底标记的乳腺谱系的表达。在腔细胞系(MCF7,SkBr3)中,培养基对转录表达的影响比在基底细胞系(MDA-MB-436)中更为明显。就补充和基础培养基而言,生长培养基的变化延迟了细胞的生长,但提高了细胞产量。结论:乳腺癌细胞分化表型标志物的表达取决于细胞生长培养基的组成,因此考虑到这种作用,应使用细胞培养作为表型研究的工具。此类研究的发现应始终谨慎解释。细胞生长培养基的配制对腔中而不是基底细胞系中表型标志物的表达具有更大的影响。含有促细胞分裂剂和较高维生素含量的培养基可提高细胞培养的细胞产量,尽管生长时间大大增加。

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