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Updated: Aug 7, 2026
Isolation of Mammary Epithelial Cells from Three-dimensional Mixed-cell Spheroid Co-culture
Published on: April 30, 2012
Bovine mammary myoepithelial cells. 2. Interactions with epithelial cells in vitro
B Zavizion1, I Politis, R C Gorewit
1Department of Animal Science, Cornell University, Ithaca, NY 14853.
Insights
Bovine mammary epithelial and myoepithelial cells interact in culture, forming structures that mimic the in vivo mammary gland organization. Myoepithelial cells organize epithelial cells, suggesting a key role in mammary gland architecture.
Area of Science:
- Veterinary Science
- Cell Biology
- Developmental Biology
Background:
- The bovine mammary gland's complex architecture is crucial for milk production.
- Understanding cell-cell interactions is vital for elucidating mammary gland development.
Purpose of the Study:
- To investigate the in vitro interactions between bovine myoepithelial and epithelial cells.
- To characterize the structural organization and behavior of these cells in coculture.
Main Methods:
- Dissociation of mammary tissue using collagenase.
- Separation of myoepithelial and epithelial cells via differential centrifugation.
- Cell culture in specific media and coculture conditions.
Main Results:
- Epithelial cell conditioned medium slightly reduced fetal myoepithelial cell proliferation.
- Myoepithelial cells extended filopodia and migrated towards epithelial cells.
- Cocultured cells formed domelike structures resembling in vivo mammary gland organization.
- Myoepithelial cells organized epithelial cells into distinct colonies.
Conclusions:
- Myoepithelial and epithelial cells exhibit specific interactions in vitro.
- These interactions lead to the formation of structures mimicking in vivo mammary gland architecture.
- Myoepithelial cells play a significant role in organizing the mammary gland's structural framework during development.
Abstract:
The objective of this study was to examine interactions of bovine myoepithelial and epithelial cells in vitro. Mammary tissue was dissociated with collagenase into myoepithelial and epithelial cells. Myoepithelial and epithelial cells were separated by differential centrifugation. Both cell types were cultured on plastic in RPMI-1640 and Iscove's Modified Dulbecco's Modified Eagle Medium supplemented with 10% horse serum and 5% fetal bovine serum. Our data revealed that conditioned medium from epithelial cells caused a small but significant reduction in proliferation of myoepithelial cells from fetal mammary glands. Myoepithelial-epithelial cell interaction in culture was characterized by myoepithelial cells with extended filopodia that could grow on top of confluent monolayers of epithelial cells, imitating the in vivo situation. In confluent monolayers of epithelial and myoepithelial cells in coculture, small domelike structures consisting of mixtures of epithelial and myoepithelial cells were observed. These structures greatly resembled the in vivo organization of the bovine mammary gland. Furthermore, myoepithelial cells were capable of migration toward individual colonies of epithelial cells or single epithelial cells. Myoepithelial cells organized epithelial cells into well-defined colonies. Myoepithelial cells may play an important role in organizing the architectural framework of the mammary gland during growth and development.
