An Evaluation of Matrix-Containing and Humanised Matrix-Free 3-Dimensional Cell Culture Systems for Studying Breast Cancer
| Autor: | Grace C, Roberts, Paul G, Morris, Marcus A, Moss, Sarah L, Maltby, Chelsea A, Palmer, Claire E, Nash, Emily, Smart, Deborah L, Holliday, Valerie, Speirs |
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| Rok vydání: | 2015 |
| Předmět: |
Cell Survival
Cell Lines Biocompatible Materials Breast Neoplasms BT474 cells Research and Analysis Methods Biochemistry Collagen Type I Epithelium Fluorescence Microscopy Animal Cells Cell Line Tumor Spheroids Cellular Breast Tumors Breast Cancer Cell Adhesion Tumor Cells Cultured Medicine and Health Sciences Humans Breast Connective Tissue Cells Microscopy Biology and Life Sciences Cancers and Neoplasms Proteins Light Microscopy Epithelial Cells Cell Biology Fibroblasts Cell Cultures Coculture Techniques Biological Tissue Oncology Connective Tissue Female Biological Cultures Cultured Fibroblasts Cellular Types Anatomy Collagens Research Article |
| Zdroj: | PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Background 3D cell cultures are emerging as more physiologically meaningful alternatives to monolayer cultures for many biological applications. They are attractive because they more closely mimic in vivo morphology, especially when co-cultured with stromal fibroblasts. Methodology/Principal Findings We compared the efficacy of 3 different 3D cell culture systems; collagen I, low attachment culture vessels and a modification of Fibrolife®, a specialised humanised cell culture medium devoid of animal-derived components, using breast cancer cell lines representative of the different molecular subtypes of breast cancer, cultured alone or with human mammary fibroblasts with a view to developing matrix-free humanised systems. 3D collagen I culture supported the growth of a range of breast cancer cell lines. By modifying the composition of Fibrolife® to epiFL, matrix-free cell culture was possible. During sequential transfer to epiFL breast cancer cells gradually detached from the flask, growing progressively as spheroids. Phenotype was stable and reversible with cells remaining actively proliferating and easily accessible throughout culture. They could also be revived from frozen stocks. To achieve co-culture with fibroblasts in epiFL required use of low attachment culture vessels instead of standard plastic as fibroblasts remained adherent in epiFL. Here, cancer cell spheroids were allowed to form before adding fibroblasts. Immunohistochemical examination showed fibroblasts scattered throughout the epithelial spheroid, not dissimilar to the relationship of tumour stroma in human breast cancer. Conclusions Because of its ease of handling, matrix-free 3D cell culture may be a useful model to study the influence of fibroblasts on breast cancer epithelial cells with use of epiFL culture medium taking this a step further towards a fully humanised 3D model. This methodology could be applied to other types of cancer cell lines, making this a versatile technique for cancer researchers wishing to use in vitro systems that better reflect cancer in vivo. |
| Databáze: | OpenAIRE |
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