In vitro analysis of breast cancer cell line tumourspheres and primary human breast epithelia mammospheres demonstrates inter- and intrasphere heterogeneity

Smart, Chanel E., Morrison, Brian J., Saunus, Jodi M., Vargas, Ana Cristina, Keith, Patricia, Reid, Lynne, Wockner, Leesa, Amiri, Marjan Askarian, Sarkar, Debina, Simpson, Peter T., Clarke, Catherine, Schmidt, Chris W., Reynolds, Brent A., Lakhani, Sunil R. and Lopez, J. Alejandro (2013) In vitro analysis of breast cancer cell line tumourspheres and primary human breast epithelia mammospheres demonstrates inter- and intrasphere heterogeneity. PloS One, 8 6: . doi:10.1371/journal.pone.0064388


Author Smart, Chanel E.
Morrison, Brian J.
Saunus, Jodi M.
Vargas, Ana Cristina
Keith, Patricia
Reid, Lynne
Wockner, Leesa
Amiri, Marjan Askarian
Sarkar, Debina
Simpson, Peter T.
Clarke, Catherine
Schmidt, Chris W.
Reynolds, Brent A.
Lakhani, Sunil R.
Lopez, J. Alejandro
Title In vitro analysis of breast cancer cell line tumourspheres and primary human breast epithelia mammospheres demonstrates inter- and intrasphere heterogeneity
Formatted title
In vitro analysis of breast cancer cell line tumourspheres and primary human breast epithelia mammospheres demonstrates inter- and intrasphere heterogeneity
Journal name PloS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2013-06-04
Year available 2013
Sub-type Article (original research)
DOI 10.1371/journal.pone.0064388
Open Access Status DOI
Volume 8
Issue 6
Total pages 15
Place of publication San Francisco, United States
Publisher Public Library of Science
Collection year 2014
Language eng
Formatted abstract
Mammosphere and breast tumoursphere culture have gained popularity as in vitro assays for propagating and analysing normal and cancer stem cells. Whether the spheres derived from different sources or parent cultures themselves are indeed single entities enriched in stem/progenitor cells compared to other culture formats has not been fully determined. We surveyed sphere-forming capacity across 26 breast cell lines, immunophenotyped spheres from six luminal- and basal-like lines by immunohistochemistry and flow cytometry and compared clonogenicity between sphere, adherent and matrigel culture formats using in vitro functional assays. Analyses revealed morphological and molecular intra- and inter sphere heterogeneity, consistent with adherent parental cell line phenotypes. Flow cytometry showed sphere culture does not universally enrich for markers previously associated with stem cell phenotypes, although we found some cell-line specific changes between sphere and adherent formats. Sphere-forming efficiency was significantly lower than adherent or matrigel clonogenicity and constant over serial passage. Surprisingly, self-renewal capacity of sphere-derived cells was similar/lower than other culture formats. We observed significant correlation between long-term-proliferating-cell symmetric division rates in sphere and adherent cultures, suggesting functional overlap between the compartments sustaining them. Experiments with normal primary human mammary epithelia, including sorted luminal (MUC1+) and basal/myoepithelial (CD10+) cells revealed distinct luminal-like, basal-like and mesenchymal entities amongst primary mammospheres. Morphological and colony-forming-cell assay data suggested mammosphere culture may enrich for a luminal progenitor phenotype, or induce reversion/relaxation of the basal/mesenchymal in vitro selection occurring with adherent culture. Overall, cell line tumourspheres and primary mammospheres are not homogenous entities enriched for stem cells, suggesting a more cautious approach to interpreting data from these assays and careful consideration of its limitations. Sphere culture may represent an alternative 3-dimensional culture system which rather than universally ‘enriching’ for stem cells, has utility as one of a suite of functional assays that provide a read-out of progenitor activity.
Keyword Mammary stem/progenitor cells
Growth-factor receptor
Stem cells
Functional-characterization
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Article # e64388

Document type: Journal Article
Sub-type: Article (original research)
Collections: UQ Centre for Clinical Research Publications
Official 2014 Collection
School of Medicine Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 11 times in Thomson Reuters Web of Science Article | Citations
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