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Title: Different culture media modulate growth, heterogeneity, and senescence in human mammary epithelial cell cultures

The ability to culture normal human mammary epithelial cells (HMEC) greatly facilitates experiments that seek to understand both normal mammary cell biology and the many differences between normal and abnormal human mammary epithelia. To maximize in vivo relevance, the primary cell culture conditions should maintain cells in states that resemble in vivo as much as possible. Towards this goal, we compared the properties of HMEC strains from two different reduction mammoplasty tissues that were grown in parallel using different media and culture conditions. Epithelial organoids were initiated into three different media: two commonly used serum-free-media, MCDB 170-type (e.g. MEGM) and WIT-P, and a low stress media, M87A. Growth, lineage heterogeneity, p16 protein expression, and population doublings to senescence were measured for each culture condition. MCDB 170 caused rapid senescence and loss of heterogeneity within 2 to 3 passages, but some cultures went through the 1 to 2 month process of selection to generate clonal finite post-selection poststasis cells. WIT-P caused impressive expansion of luminal cells in 2nd passage followed by their near complete disappearance by passage 4 and senescence shortly thereafter. M87A supported as much as twice the number of population doublings compared to either serumfree medium, and luminal andmore » myoepithelial cells were present for as many as 8 passages. Thus, of the three media compared, WIT-P and MCDB 170 imposed rapid senescence and loss of lineage heterogeneity, phenotypes consistent with cells maintained in high-stress conditions, while M87A supported cultures that maintained multiple lineages and robust growth for up to 60 population doublings. In conjunction with previous studies examining the molecular properties of cultures grown in these media, we conclude that M87A medium is most able to support long-term culture of multiple lineages similar to in vivo conditions, thereby facilitating investigations of normal HMEC biology relevant to the mammary gland in situ.« less
Authors:
; ; ; ; ; ORCiD logo ;
Publication Date:
Grant/Contract Number:
AC02-05CH11231
Type:
Accepted Manuscript
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 13; Journal Issue: 10; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC)
Country of Publication:
United States
Language:
English
OSTI Identifier:
1494089

Lee, Jonathan K., Bloom, Jessica, Zubeldia-Plazaola, Arantzazu, Garbe, James C., Stampfer, Martha R., LaBarge, Mark A., and Chalmers, Jeffrey. Different culture media modulate growth, heterogeneity, and senescence in human mammary epithelial cell cultures. United States: N. p., Web. doi:10.1371/journal.pone.0204645.
Lee, Jonathan K., Bloom, Jessica, Zubeldia-Plazaola, Arantzazu, Garbe, James C., Stampfer, Martha R., LaBarge, Mark A., & Chalmers, Jeffrey. Different culture media modulate growth, heterogeneity, and senescence in human mammary epithelial cell cultures. United States. doi:10.1371/journal.pone.0204645.
Lee, Jonathan K., Bloom, Jessica, Zubeldia-Plazaola, Arantzazu, Garbe, James C., Stampfer, Martha R., LaBarge, Mark A., and Chalmers, Jeffrey. 2018. "Different culture media modulate growth, heterogeneity, and senescence in human mammary epithelial cell cultures". United States. doi:10.1371/journal.pone.0204645.
@article{osti_1494089,
title = {Different culture media modulate growth, heterogeneity, and senescence in human mammary epithelial cell cultures},
author = {Lee, Jonathan K. and Bloom, Jessica and Zubeldia-Plazaola, Arantzazu and Garbe, James C. and Stampfer, Martha R. and LaBarge, Mark A. and Chalmers, Jeffrey},
abstractNote = {The ability to culture normal human mammary epithelial cells (HMEC) greatly facilitates experiments that seek to understand both normal mammary cell biology and the many differences between normal and abnormal human mammary epithelia. To maximize in vivo relevance, the primary cell culture conditions should maintain cells in states that resemble in vivo as much as possible. Towards this goal, we compared the properties of HMEC strains from two different reduction mammoplasty tissues that were grown in parallel using different media and culture conditions. Epithelial organoids were initiated into three different media: two commonly used serum-free-media, MCDB 170-type (e.g. MEGM) and WIT-P, and a low stress media, M87A. Growth, lineage heterogeneity, p16 protein expression, and population doublings to senescence were measured for each culture condition. MCDB 170 caused rapid senescence and loss of heterogeneity within 2 to 3 passages, but some cultures went through the 1 to 2 month process of selection to generate clonal finite post-selection poststasis cells. WIT-P caused impressive expansion of luminal cells in 2nd passage followed by their near complete disappearance by passage 4 and senescence shortly thereafter. M87A supported as much as twice the number of population doublings compared to either serumfree medium, and luminal and myoepithelial cells were present for as many as 8 passages. Thus, of the three media compared, WIT-P and MCDB 170 imposed rapid senescence and loss of lineage heterogeneity, phenotypes consistent with cells maintained in high-stress conditions, while M87A supported cultures that maintained multiple lineages and robust growth for up to 60 population doublings. In conjunction with previous studies examining the molecular properties of cultures grown in these media, we conclude that M87A medium is most able to support long-term culture of multiple lineages similar to in vivo conditions, thereby facilitating investigations of normal HMEC biology relevant to the mammary gland in situ.},
doi = {10.1371/journal.pone.0204645},
journal = {PLoS ONE},
number = 10,
volume = 13,
place = {United States},
year = {2018},
month = {10}
}

Works referenced in this record:

Expression of the telomerase catalytic subunit, hTERT, induces resistance to transforming growth factor ? growth inhibition in p16INK4A(?) human mammary epithelial cells
journal, April 2001
  • Stampfer, M. R.; Garbe, J.; Levine, G.
  • Proceedings of the National Academy of Sciences, Vol. 98, Issue 8, p. 4498-4503
  • DOI: 10.1073/pnas.071483998