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

Autor:	Martha R. Stampfer, James C. Garbe, Jessica Bloom, Jonathan K. Lee, Mark A. LaBarge, Arantzazu Zubeldia-Plazaola
Přispěvatelé:	Chalmers, Jeffrey
Jazyk:	angličtina
Rok vydání:	2018
Předmět:	0301 basic medicine Mammaplasty Cellular differentiation Mammary gland lcsh:Medicine Epithelium Spectrum Analysis Techniques Animal Cells Medicine and Health Sciences Organ Cultures lcsh:Science Cells Cultured Cellular Senescence Staining education.field_of_study Cultured Multidisciplinary Ecology medicine.diagnostic_test Cell Staining Cell Differentiation Middle Aged Flow Cytometry Mammary Glands Phenotype Cell biology Organoids Laboratory Equipment medicine.anatomical_structure Spectrophotometry Engineering and Technology Female Biological Cultures Cytophotometry Cellular Types Anatomy Human Research Article Adult Senescence Ecological Metrics General Science & Technology Cells 1.1 Normal biological development and functioning Population Equipment Biology Research and Analysis Methods Flow cytometry Young Adult 03 medical and health sciences Underpinning research MD Multidisciplinary medicine Humans Cell Lineage Mammary Glands Human education Ecology and Environmental Sciences lcsh:R Myoepithelial cell Biology and Life Sciences Species Diversity Epithelial Cells Cell Biology Cell Cultures Culture Media Biological Tissue 030104 developmental biology Specimen Preparation and Treatment Cell culture lcsh:Q Developmental Biology
Zdroj:	PLoS ONE, Vol 13, Iss 10, p e0204645 (2018) PLoS ONE PloS one, vol 13, iss 10 Lee, JK; Bloom, J; Zubeldia-Plazaola, A; Garbe, JC; Stampfer, MR; & LaBarge, MA. (2018). Different culture media modulate growth, heterogeneity, and senescence in human mammary epithelial cell cultures. PLoS ONE, 13(10), e0204645. doi: 10.1371/journal.pone.0204645. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/9bt759gm
ISSN:	1932-6203
Popis:	© 2018 Lee et al. 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.
Databáze:	OpenAIRE
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