Sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypes
Abstract Background A challenge in human mammary epithelial cell (HMEC) culture is sustaining the representation of competing luminal, myoepithelial, and progenitor lineages over time. As cells replicate in culture, myoepithelial cells come to dominate the composition of the culture with serial pass...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-01-01
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| Series: | Breast Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13058-022-01595-z |
| _version_ | 1797945680302440448 |
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| author | Michael E. Todhunter Masaru Miyano Eric G. Carlson Stefan Hinz Mark A. LaBarge |
| author_facet | Michael E. Todhunter Masaru Miyano Eric G. Carlson Stefan Hinz Mark A. LaBarge |
| author_sort | Michael E. Todhunter |
| collection | DOAJ |
| description | Abstract Background A challenge in human mammary epithelial cell (HMEC) culture is sustaining the representation of competing luminal, myoepithelial, and progenitor lineages over time. As cells replicate in culture, myoepithelial cells come to dominate the composition of the culture with serial passaging. This drift in composition presents a challenge for studying luminal and progenitor cells, which are prospective cells of origin for most breast cancer subtypes. Methods We demonstrate the use of postconfluent culture on HMECs. Postconfluent culture entails culturing HMECs for 2–5 weeks without passaging but maintaining frequent feedings in low-stress M87A culture medium. In contrast, standard HMEC culture entails enzymatic subculturing every 3–5 days to maintain subconfluent density. Results When compared to standard HMEC culture, postconfluent culture yields increased proportions of luminal cells and c-Kit+ progenitor cells. Postconfluent cultures develop a distinct multilayered morphology with individual cells showing decreased physical deformability as compared to cells in standard culture. Gene expression analysis of postconfluent cells shows increased expression of lineage-specific markers and extracellular matrix components. Conclusions Postconfluent culture is a novel, useful strategy for altering the lineage composition of HMECs, by increasing the proportional representation of luminal and progenitor cells. We speculate that postconfluent culture creates a microenvironment with cellular composition closer to the physiological state and eases the isolation of scarce cell subtypes. As such, postconfluent culture is a valuable tool for researchers using HMECs for breast cancer research. |
| first_indexed | 2024-04-10T20:58:56Z |
| format | Article |
| id | doaj.art-7da9d03f9ca04167a9df2a43dd1a680d |
| institution | Directory Open Access Journal |
| issn | 1465-542X |
| language | English |
| last_indexed | 2024-04-10T20:58:56Z |
| publishDate | 2023-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Breast Cancer Research |
| spelling | doaj.art-7da9d03f9ca04167a9df2a43dd1a680d2023-01-22T12:28:55ZengBMCBreast Cancer Research1465-542X2023-01-0125111510.1186/s13058-022-01595-zSustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypesMichael E. Todhunter0Masaru Miyano1Eric G. Carlson2Stefan Hinz3Mark A. LaBarge4Department of Population Sciences, Beckman Research Institute at City of HopeDepartment of Population Sciences, Beckman Research Institute at City of HopeDepartment of Population Sciences, Beckman Research Institute at City of HopeDepartment of Population Sciences, Beckman Research Institute at City of HopeDepartment of Population Sciences, Beckman Research Institute at City of HopeAbstract Background A challenge in human mammary epithelial cell (HMEC) culture is sustaining the representation of competing luminal, myoepithelial, and progenitor lineages over time. As cells replicate in culture, myoepithelial cells come to dominate the composition of the culture with serial passaging. This drift in composition presents a challenge for studying luminal and progenitor cells, which are prospective cells of origin for most breast cancer subtypes. Methods We demonstrate the use of postconfluent culture on HMECs. Postconfluent culture entails culturing HMECs for 2–5 weeks without passaging but maintaining frequent feedings in low-stress M87A culture medium. In contrast, standard HMEC culture entails enzymatic subculturing every 3–5 days to maintain subconfluent density. Results When compared to standard HMEC culture, postconfluent culture yields increased proportions of luminal cells and c-Kit+ progenitor cells. Postconfluent cultures develop a distinct multilayered morphology with individual cells showing decreased physical deformability as compared to cells in standard culture. Gene expression analysis of postconfluent cells shows increased expression of lineage-specific markers and extracellular matrix components. Conclusions Postconfluent culture is a novel, useful strategy for altering the lineage composition of HMECs, by increasing the proportional representation of luminal and progenitor cells. We speculate that postconfluent culture creates a microenvironment with cellular composition closer to the physiological state and eases the isolation of scarce cell subtypes. As such, postconfluent culture is a valuable tool for researchers using HMECs for breast cancer research.https://doi.org/10.1186/s13058-022-01595-zHuman mammary epithelial cultureLuminalMyoepithelialc-KitPostconfluentMultilayered |
| spellingShingle | Michael E. Todhunter Masaru Miyano Eric G. Carlson Stefan Hinz Mark A. LaBarge Sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypes Breast Cancer Research Human mammary epithelial culture Luminal Myoepithelial c-Kit Postconfluent Multilayered |
| title | Sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypes |
| title_full | Sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypes |
| title_fullStr | Sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypes |
| title_full_unstemmed | Sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypes |
| title_short | Sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c-Kit+ subtypes |
| title_sort | sustained postconfluent culture of human mammary epithelial cells enriches for luminal and c kit subtypes |
| topic | Human mammary epithelial culture Luminal Myoepithelial c-Kit Postconfluent Multilayered |
| url | https://doi.org/10.1186/s13058-022-01595-z |
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