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Breast Cancer Research
Published in: Breast Cancer Research 3/2014

Open Access 01-06-2014 | Research article

Mesenchymal precursor cells maintain the differentiation and proliferation potentials of breast epithelial cells

Authors: Stephan Duss, Heike Brinkhaus, Adrian Britschgi, Erik Cabuy, Daniel M Frey, Dirk J Schaefer, Mohamed Bentires-Alj

Published in: Breast Cancer Research | Issue 3/2014

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Abstract

Introduction

Stromal-epithelial interactions play a fundamental role in tissue homeostasis, controlling cell proliferation and differentiation. Not surprisingly, aberrant stromal-epithelial interactions contribute to malignancies. Studies of the cellular and molecular mechanisms underlying these interactions require ex vivo experimental model systems that recapitulate the complexity of human tissue without compromising the differentiation and proliferation potentials of human primary cells.

Methods

We isolated and characterized human breast epithelial and mesenchymal precursors from reduction mammoplasty tissue and tagged them with lentiviral vectors. We assembled heterotypic co-cultures and compared mesenchymal and epithelial cells to cells in corresponding monocultures by analyzing growth, differentiation potentials, and gene expression profiles.

Results

We show that heterotypic culture of non-immortalized human primary breast epithelial and mesenchymal precursors maintains their proliferation and differentiation potentials and constrains their growth. We further describe the gene expression profiles of stromal and epithelial cells in co-cultures and monocultures and show increased expression of the tumor growth factor beta (TGFβ) family member inhibin beta A (INHBA) in mesenchymal cells grown as co-cultures compared with monocultures. Notably, overexpression of INHBA in mesenchymal cells increases colony formation potential of epithelial cells, suggesting that it contributes to the dynamic reciprocity between breast mesenchymal and epithelial cells.

Conclusions

The described heterotypic co-culture system will prove useful for further characterization of the molecular mechanisms mediating interactions between human normal or neoplastic breast epithelial cells and the stroma, and will provide a framework to test the relevance of the ever-increasing number of oncogenomic alterations identified in human breast cancer.
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Literature
1.
Ince TA, Richardson AL, Bell GW, Saitoh M, Godar S, Karnoub AE, Iglehart JD, Weinberg RA: Transformation of different human breast epithelial cell types leads to distinct tumor phenotypes. Cancer Cell. 2007, 12: 160-170. 10.1016/j.ccr.2007.06.013.CrossRefPubMed
2.
Lim E, Vaillant F, Wu D, Forrest NC, Pal B, Hart AH, Asselin-Labat ML, Gyorki DE, Ward T, Partanen A, Feleppa F, Huschtscha LI, Thorne H, Fox SB, Yan M, French JD, Brown MA, Smyth GK, Visvader JE, Lindeman GJ, kConFab: Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers. Nat Med. 2009, 15: 907-913. 10.1038/nm.2000.CrossRefPubMed
3.
4.
Tlsty TD, Coussens LM: Tumor stroma and regulation of cancer development. Annu Rev Pathol. 2006, 1: 119-150. 10.1146/annurev.pathol.1.110304.100224.CrossRefPubMed
5.
Egeblad M, Nakasone ES, Werb Z: Tumors as organs: complex tissues that interface with the entire organism. Dev Cell. 2010, 18: 884-901. 10.1016/j.devcel.2010.05.012.CrossRefPubMedPubMedCentral
6.
7.
Laurence DJ, Monaghan P, Gusterson BA: The development of the normal human breast. Oxf Rev Reprod Biol. 1991, 13: 149-174.PubMed
8.
9.
Smith CA, Monaghan P, Neville AM: Basal clear cells of the normal human breast. Virchows Arch A Pathol Anat Histopathol. 1984, 402: 319-329. 10.1007/BF00695085.CrossRefPubMed
10.
Stingl J, Eaves CJ, Kuusk U, Emerman JT: Phenotypic and functional characterization in vitro of a multipotent epithelial cell present in the normal adult human breast. Differentiation. 1998, 63: 201-213. 10.1111/j.1432-0436.1998.00201.x.CrossRefPubMed
11.
Stingl J, Eaves CJ, Zandieh I, Emerman JT: Characterization of bipotent mammary epithelial progenitor cells in normal adult human breast tissue. Breast Cancer Res Treat. 2001, 67: 93-109. 10.1023/A:1010615124301.CrossRefPubMed
12.
Li L, Xie T: Stem cell niche: structure and function. Annu Rev Cell Dev Biol. 2005, 21: 605-631. 10.1146/annurev.cellbio.21.012704.131525.CrossRefPubMed
13.
Schofield R: The relationship between the spleen colony-forming cell and the haemopoietic stem cell. Blood Cells. 1978, 4: 7-25.PubMed
14.
15.
Wilson A, Trumpp A: Bone-marrow haematopoietic-stem-cell niches. Nat Rev Immunol. 2006, 6: 93-106. 10.1038/nri1779.CrossRefPubMed
16.
Zhang J, Niu C, Ye L, Huang H, He X, Tong WG, Ross J, Haug J, Johnson T, Feng JQ, Harris S, Wiedemann LM, Mishina Y, Li L: Identification of the haematopoietic stem cell niche and control of the niche size. Nature. 2003, 425: 836-841. 10.1038/nature02041.CrossRefPubMed
17.
Chepko G, Smith GH: Three division-competent, structurally-distinct cell populations contribute to murine mammary epithelial renewal. Tissue Cell. 1997, 29: 239-253. 10.1016/S0040-8166(97)80024-9.CrossRefPubMed
18.
Brisken C, Duss S: Stem cells and the stem cell niche in the breast: an integrated hormonal and developmental perspective. Stem Cell Rev. 2007, 3: 147-156. 10.1007/s12015-007-0019-1.CrossRefPubMed
19.
20.
Mendez-Ferrer S, Michurina TV, Ferraro F, Mazloom AR, Macarthur BD, Lira SA, Scadden DT, Ma'ayan A, Enikolopov GN, Frenette PS: Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature. 2010, 466: 829-834. 10.1038/nature09262.CrossRefPubMedPubMedCentral
21.
Friedenstein AJ, Petrakova KV, Kurolesova AI, Frolova GP: Heterotopic of bone marrow, Analysis of precursor cells for osteogenic and hematopoietic tissues. Transplantation. 1968, 6: 230-247. 10.1097/00007890-196803000-00009.CrossRefPubMed
22.
Valtieri M, Sorrentino A: The mesenchymal stromal cell contribution to homeostasis. J Cell Physiol. 2008, 217: 296-300. 10.1002/jcp.21521.CrossRefPubMed
23.
Beyer Nardi N, da Silva ML: Mesenchymal stem cells: isolation, in vitro expansion and characterization. Handb Exp Pharmacol. 2006, 174: 249-282.CrossRef
24.
Dennis JE, Charbord P: Origin and differentiation of human and murine stroma. Stem Cells. 2002, 20: 205-214. 10.1634/stemcells.20-3-205.CrossRefPubMed
25.
Liu ZJ, Zhuge Y, Velazquez OC: Trafficking and differentiation of mesenchymal stem cells. J Cell Biochem. 2009, 106: 984-991. 10.1002/jcb.22091.CrossRefPubMed
26.
Cunha GR, Young P, Christov K, Guzman R, Nandi S, Talamantes F, Thordarson G: Mammary phenotypic expression induced in epidermal cells by embryonic mammary mesenchyme. Acta Anat (Basel). 1995, 152: 195-204. 10.1159/000147698.CrossRef
27.
Heuberger B, Fitzka I, Wasner G, Kratochwil K: Induction of androgen receptor formation by epithelium-mesenchyme interaction in embryonic mouse mammary gland. Proc Natl Acad Sci U S A. 1982, 79: 2957-2961. 10.1073/pnas.79.9.2957.CrossRefPubMedPubMedCentral
28.
Robinson GW, Karpf AB, Kratochwil K: Regulation of mammary gland development by tissue interaction. J Mammary Gland Biol Neoplasia. 1999, 4: 9-19. 10.1023/A:1018748418447.CrossRefPubMed
29.
van Genderen C, Okamura RM, Farinas I, Quo RG, Parslow TG, Bruhn L, Grosschedl R: Development of several organs that require inductive epithelial-mesenchymal interactions is impaired in LEF-1-deficient mice. Genes Dev. 1994, 8: 2691-2703. 10.1101/gad.8.22.2691.CrossRefPubMed
30.
Hughes ES: The Development of the Mammary Gland: Arris and Gale Lecture, delivered at the Royal College of Surgeons of England on 25th October, 1949. Ann R Coll Surg Engl. 1950, 6: 99-119.PubMedPubMedCentral
31.
Ellis MJ, Ding L, Shen D, Luo J, Suman VJ, Wallis JW, Van Tine BA, Hoog J, Goiffon RJ, Goldstein TC, Ng S, Lin L, Crowder R, Snider J, Ballman K, Weber J, Chen K, Koboldt DC, Kandoth C, Schierding WS, McMichael JF, Miller CA, Lu C, Harris CC, McLellan MD, Wendl MC, DeSchryver K, Allred DC, Esserman L, Unzeitig G, et al: Whole-genome analysis informs breast cancer response to aromatase inhibition. Nature. 2012, 486: 353-360.PubMedPubMedCentral
32.
Banerji S, Cibulskis K, Rangel-Escareno C, Brown KK, Carter SL, Frederick AM, Lawrence MS, Sivachenko AY, Sougnez C, Zou L, Cortes ML, Fernandez-Lopez JC, Peng S, Ardlie KG, Auclair D, Bautista-Piña V, Duke F, Francis J, Jung J, Maffuz-Aziz A, Onofrio RC, Parkin M, Pho NH, Quintanar-Jurado V, Ramos AH, Rebollar-Vega R, Rodriguez-Cuevas S, Romero-Cordoba SL, Schumacher SE, Stransky N, et al: Sequence analysis of mutations and translocations across breast cancer subtypes. Nature. 2012, 486: 405-409. 10.1038/nature11154.CrossRefPubMedPubMedCentral
33.
Curtis C, Shah SP, Chin SF, Turashvili G, Rueda OM, Dunning MJ, Speed D, Lynch AG, Samarajiwa S, Yuan Y, Gräf S, Ha G, Haffari G, Bashashati A, Russell R, McKinney S, Langerød A, Green A, Provenzano E, Wishart G, Pinder S, Watson P, Markowetz F, Murphy L, Ellis I, Purushotham A, Børresen-Dale AL, Brenton JD, Tavaré S, METABRIC Group, et al: The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature. 2012, 486: 346-352.PubMedPubMedCentral
34.
Nelson CM, Bissell MJ: Modeling dynamic reciprocity: engineering three-dimensional culture models of breast architecture, function, and neoplastic transformation. Semin Cancer Biol. 2005, 15: 342-352. 10.1016/j.semcancer.2005.05.001.CrossRefPubMedPubMedCentral
35.
Weigelt B, Bissell MJ: Unraveling the microenvironmental influences on the normal mammary gland and breast cancer. Semin Cancer Biol. 2008, 18: 311-321. 10.1016/j.semcancer.2008.03.013.CrossRefPubMedPubMedCentral
36.
Park JR, Jung JW, Lee YS, Kang KS: The roles of Wnt antagonists Dkk1 and sFRP4 during adipogenesis of human adipose tissue-derived mesenchymal stem cells. Cell Prolif. 2008, 41: 859-874. 10.1111/j.1365-2184.2008.00565.x.CrossRefPubMed
37.
Stampfer M, Hallowes RC, Hackett AJ: Growth of normal human mammary cells in culture. In Vitro. 1980, 16: 415-425. 10.1007/BF02618365.CrossRefPubMed
38.
Stingl J, Emerman JT, Eaves CJ: Enzymatic dissociation and culture of normal human mammary tissue to detect progenitor activity. Methods Mol Biol. 2005, 290: 249-263.PubMed
39.
Lin TM, Tsai JL, Lin SD, Lai CS, Chang CC: Accelerated growth and prolonged lifespan of adipose tissue-derived human mesenchymal stem cells in a medium using reduced calcium and antioxidants. Stem Cells Dev. 2005, 14: 92-102. 10.1089/scd.2005.14.92.CrossRefPubMed
40.
Dontu G, Abdallah WM, Foley JM, Jackson KW, Clarke MF, Kawamura MJ, Wicha MS: In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev. 2003, 17: 1253-1270. 10.1101/gad.1061803.CrossRefPubMedPubMedCentral
41.
Duss S, Andre S, Nicoulaz AL, Fiche M, Bonnefoi H, Brisken C, Iggo RD: An oestrogen-dependent model of breast cancer created by transformation of normal human mammary epithelial cells. Breast Cancer Res. 2007, 9: R38-10.1186/bcr1734.CrossRefPubMedPubMedCentral
42.
Nagoshi E, Saini C, Bauer C, Laroche T, Naef F, Schibler U: Circadian gene expression in individual fibroblasts: cell-autonomous and self-sustained oscillators pass time to daughter cells. Cell. 2004, 119: 693-705. 10.1016/j.cell.2004.11.015.CrossRefPubMed
43.
Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L: A third-generation lentivirus vector with a conditional packaging system. J Virol. 1998, 72: 8463-8471.PubMedPubMedCentral
44.
45.
46.
47.
Gene Set Enrichment Analysis (GSEA), computational method that determines whether an a priori defined set of genes shows statistically significant, concordant differences between two biological states. http://​www.​broadinstitute.​org/​gsea,
48.
49.
Edgar R, Domrachev M, Lash AE: Gene Expression Omnibus: NCBI gene expression and hybridization array data repository. Nucleic Acids Res. 2002, 30: 207-210. 10.1093/nar/30.1.207.CrossRefPubMedPubMedCentral
50.
51.
52.
Toussaint O, Weemaels G, Debacq-Chainiaux F, Scharffetter-Kochanek K, Wlaschek M: Artefactual effects of oxygen on cell culture models of cellular senescence and stem cell biology. J Cell Physiol. 2011, 226: 315-321. 10.1002/jcp.22416.CrossRefPubMed
53.
Wright WE, Shay JW: Inexpensive low-oxygen incubators. Nat Protoc. 2006, 1: 2088-2090. 10.1038/nprot.2006.374.CrossRefPubMed
54.
Pandis N, Heim S, Bardi G, Limon J, Mandahl N, Mitelman F: Improved technique for short-term culture and cytogenetic analysis of human breast cancer. Genes Chromosomes Cancer. 1992, 5: 14-20. 10.1002/gcc.2870050103.CrossRefPubMed
55.
Steele JG, Dalton BA, Johnson G, Underwood PA: Adsorption of fibronectin and vitronectin onto Primaria and tissue culture polystyrene and relationship to the mechanism of initial attachment of human vein endothelial cells and BHK-21 fibroblasts. Biomaterials. 1995, 16: 1057-1067. 10.1016/0142-9612(95)98901-P.CrossRefPubMed
56.
Villadsen R, Fridriksdottir AJ, Ronnov-Jessen L, Gudjonsson T, Rank F, LaBarge MA, Bissell MJ, Petersen OW: Evidence for a stem cell hierarchy in the adult human breast. J Cell Biol. 2007, 177: 87-101. 10.1083/jcb.200611114.CrossRefPubMedPubMedCentral
57.
Shahdadfar A, Fronsdal K, Haug T, Reinholt FP, Brinchmann JE: In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression, and transcriptome stability. Stem Cells. 2005, 23: 1357-1366. 10.1634/stemcells.2005-0094.CrossRefPubMed
58.
Petersen OW, Ronnov-Jessen L, Howlett AR, Bissell MJ: Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. Proc Natl Acad Sci U S A. 1992, 89: 9064-9068. 10.1073/pnas.89.19.9064.CrossRefPubMedPubMedCentral
59.
Debnath J, Mills KR, Collins NL, Reginato MJ, Muthuswamy SK, Brugge JS: The role of apoptosis in creating and maintaining luminal space within normal and oncogene-expressing mammary acini. Cell. 2002, 111: 29-40. 10.1016/S0092-8674(02)01001-2.CrossRefPubMed
60.
Desmedt C, Majjaj S, Kheddoumi N, Singhal SK, Haibe-Kains B, El Ouriaghli F, Chaboteaux C, Michiels S, Lallemand F, Journe F, Duvillier H, Loi S, Quackenbush J, Dekoninck S, Blanpain C, Lagneaux L, Houhou N, Delorenzi M, Larsimont D, Piccart M, Sotiriou C: Characterization and clinical evaluation of CD10+ stroma cells in the breast cancer microenvironment. Clin Cancer Res. 2012, 18: 1004-1014. 10.1158/1078-0432.CCR-11-0383.CrossRefPubMedPubMedCentral
61.
Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, Paulovich A, Pomeroy SL, Golub TR, Lander ES, Mesirov JP: Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A. 2005, 102: 15545-15550. 10.1073/pnas.0506580102.CrossRefPubMedPubMedCentral
62.
Subramanian A, Kuehn H, Gould J, Tamayo P, Mesirov JP: GSEA-P: a desktop application for Gene Set Enrichment Analysis. Bioinformatics. 2007, 23: 3251-3253. 10.1093/bioinformatics/btm369.CrossRefPubMed
63.
Allinen M, Beroukhim R, Cai L, Brennan C, Lahti-Domenici J, Huang H, Porter D, Hu M, Chin L, Richardson A, Schnitt S, Sellers WR, Polyak K: Molecular characterization of the tumor microenvironment in breast cancer. Cancer Cell. 2004, 6: 17-32. 10.1016/j.ccr.2004.06.010.CrossRefPubMed
64.
Lim E, Wu D, Pal B, Bouras T, Asselin-Labat ML, Vaillant F, Yagita H, Lindeman GJ, Smyth GK, Visvader JE: Transcriptome analyses of mouse and human mammary cell subpopulations reveal multiple conserved genes and pathways. Breast Cancer Res. 2010, 12: R21-10.1186/bcr2560.CrossRefPubMedPubMedCentral
65.
Huang TS, Hsieh JY, Wu YH, Jen CH, Tsuang YH, Chiou SH, Partanen J, Anderson H, Jaatinen T, Yu YH, Wang HW: Functional network reconstruction reveals somatic stemness genetic maps and dedifferentiation-like transcriptome reprogramming induced by GATA2. Stem Cells. 2008, 26: 1186-1201. 10.1634/stemcells.2007-0821.CrossRefPubMed
66.
Pedemonte E, Benvenuto F, Casazza S, Mancardi G, Oksenberg JR, Uccelli A, Baranzini SE: The molecular signature of therapeutic mesenchymal stem cells exposes the architecture of the hematopoietic stem cell niche synapse. BMC Genomics. 2007, 8: 65-10.1186/1471-2164-8-65.CrossRefPubMedPubMedCentral
67.
Bradley TR, Hodgson GS, Rosendaal M: The effect of oxygen tension on haemopoietic and fibroblast cell proliferation in vitro. J Cell Physiol. 1978, 97: 517-522. 10.1002/jcp.1040970327.CrossRefPubMed
68.
Brewer GJ, Cotman CW: Survival and growth of hippocampal neurons in defined medium at low density: advantages of a sandwich culture technique or low oxygen. Brain Res. 1989, 494: 65-74. 10.1016/0006-8993(89)90144-3.CrossRefPubMed
69.
Eliasson P, Jonsson JI: The hematopoietic stem cell niche: low in oxygen but a nice place to be. J Cell Physiol. 2010, 222: 17-22. 10.1002/jcp.21908.CrossRefPubMed
70.
Mohyeldin A, Garzon-Muvdi T, Quinones-Hinojosa A: Oxygen in stem cell biology: a critical component of the stem cell niche. Cell Stem Cell. 2010, 7: 150-161. 10.1016/j.stem.2010.07.007.CrossRefPubMed
71.
Wang X, Kaplan DL: Hormone-responsive 3D multicellular culture model of human breast tissue. Biomaterials. 2012, 33: 3411-3420. 10.1016/j.biomaterials.2012.01.011.CrossRefPubMedPubMedCentral
72.
Chepko G, Dickson RB: Ultrastructure of the putative stem cell niche in rat mammary epithelium. Tissue Cell. 2003, 35: 83-93. 10.1016/S0040-8166(02)00107-6.CrossRefPubMed
73.
Watt FM: The stem cell compartment in human interfollicular epidermis. J Dermatol Sci. 2002, 28: 173-180. 10.1016/S0923-1811(02)00003-8.CrossRefPubMed
74.
Yen TH, Wright NA: The gastrointestinal tract stem cell niche. Stem Cell Rev. 2006, 2: 203-212. 10.1007/s12015-006-0048-1.CrossRefPubMed
75.
Metadata
Title
Mesenchymal precursor cells maintain the differentiation and proliferation potentials of breast epithelial cells
Authors
Stephan Duss
Heike Brinkhaus
Adrian Britschgi
Erik Cabuy
Daniel M Frey
Dirk J Schaefer
Mohamed Bentires-Alj
Publication date
01-06-2014
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 3/2014
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr3673

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