Establishment of a 3D cell culture model of primary bovine mammary epithelial cells extracted from fresh milk

被引:0
作者
Maria Hillreiner
Nadine I. Müller
Heiner M. Koch
Christiane Schmautz
Bernhard Küster
Michael W. Pfaffl
Heike Kliem
机构
[1] Technische Universität München,Chair of Animal Physiology and Immunology
[2] Technische Universität München,Chair of Proteomics and Bioanalytics
[3] Technische Universität München,Bavarian Biomolecular Mass Spectrometry Center
来源
In Vitro Cellular & Developmental Biology - Animal | 2017年 / 53卷
关键词
Primary bovine mammary epithelial cells; 3D cell culture; Lactogenesis; Proteomics; RT-qPCR; Extracellular matrix;
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学科分类号
摘要
For the investigation of molecular processes underlying diseases of the bovine mammary gland, primary bovine mammary epithelial cells (pbMEC) are used. They are known to contribute to the innate immune system of the bovine mammary gland. The functionality of pbMEC depends on the maintenance of in vivo characteristics. So far, the optimization of pbMEC culture conditions was intended in a variety of experiments. For this purpose, most of the studies used stable cell lines or primary cells obtained from udder biopsies of slaughtered animals. By contrast, within our study, pbMEC of healthy and first lactating Brown Swiss cows were non-invasively isolated from fresh milk. The non-invasively isolated pbMEC were cultivated on the extracellular matrix-like scaffold Matrigel®. Further, they were challenged with different compositions of proliferation media, containing lactogenic hormones and/or the essential amino acid l-lysine. Changes in expression levels of genes coding for milk proteins and for components of the janus kinase/signal transducers and activators of transcription (JAK-STAT) and mTOR pathways were analyzed by RT-qPCR. The secreted proteins were analyzed by LC-MS/MS measurements. We showed for the first time the establishment of a physiologically functional 3D cell culture model of pbMEC isolated from fresh milk. This represents a primary cell culture model system, based on non-invasive cell collection, that can be used to unravel physiological processes in an unbiased manner.
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页码:706 / 720
页数:14
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