AI Article Synopsis
- Developed a 3D organotypic culture system for primary human mammary epithelial cells (HMECs) using free-floating collagen type I gels.
- Luminal cells form multicellular spheres, while basal/myoepithelial cells create complex structures mimicking terminal ductal lobular units (TDLUs).
- The culture supports analysis of morphology, protein expression, and gene analysis, making it ideal for researching branching morphogenesis, regeneration, and environmental effects on HMECs.
Article Abstract
We have developed a three-dimensional organotypic culture system for primary human mammary epithelial cells (HMECs) in which the cells are cultured in free floating collagen type I gels. In this assay, luminal cells predominantly form multicellular spheres, while basal/myoepithelial cells form complex branched structures resembling terminal ductal lobular units (TDLUs), the functional units of the human mammary gland in situ. The TDLU-like organoids can be cultured for at least 3 weeks and can then be passaged multiple times. Subsequently, collagen gels can be stained with carmine or by immunofluorescence to allow for the analysis of morphology, protein expression and polarization, and to facilitate quantification of structures. In addition, structures can be isolated for gene expression analysis. In summary, this technique is suitable for studying branching morphogenesis, regeneration, and differentiation of HMECs as well as their dependence on the physical environment.
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| http://dx.doi.org/10.1007/978-1-4939-7021-6_9 | DOI Listing |
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