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PMID: 12756227 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells.

Genes & development ·Vol. 17 ·No. 10 ·2003-05-15 ·Pages 1253-70

Dontu G, Abdallah WM, Foley JM, Jackson KW, Clarke MF, Kawamura MJ, Wicha MS

Abstract

Although the existence of mammary stem cells has been suggested by serial transplantation studies in mice, their identification has been hindered by the lack of specific surface markers, and by the absence of suitable in vitro assays for testing stem cell properties: self-renewal and ability to generate differentiated progeny. We have developed an in vitro cultivation system that allows for propagation of human mammary epithelial cells (HMECs) in an undifferentiated state, based on their ability to proliferate in suspension, as nonadherent mammospheres. We demonstrate that nonadherent mammospheres are enriched in early progenitor/stem cells and able to differentiate along all three mammary epithelial lineages and to clonally generate complex functional structures in reconstituted 3D culture systems. Gene expression analysis of cells isolated from nonadherent mammospheres revealed overlapping genetic programs with other stem and progenitor cells and identified new markers that may be useful in the identification of mammary stem cells. The isolation and characterization of these stem cells should help elucidate the molecular pathways that govern normal mammary development and carcinogenesis.

MeSH Terms
Benzimidazoles Biological Assay Breast/cytology,growth & development Cell Division/physiology Cell Line Epithelium/growth & development Female Fluorescent Dyes Gene Expression Profiling Gene Expression Regulation Humans In Vitro Techniques Staining and Labeling Stem Cells/physiology Transcription, Genetic
Chemicals
Benzimidazoles Fluorescent Dyes bisbenzimide ethoxide trihydrochloride
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Dontu Gabriela
Department of Internal Medicine, Hematology-Oncology, Comprehensive Cancer Center, University of Michigan, Ann Arbor, Michigan 48109, USA. gdontu@umich.edu
Abdallah Wissam M
Foley Jessica M
Jackson Kyle W
Clarke Michael F
Kawamura Mari J
Wicha Max S
References (52)
52 references, click to expand
  1. Sca-1(pos) cells in the mouse mammary gland represent an enriched progenitor cell population.
    Dev Biol. 2002 May 1;245(1):42-56 PMID: 11969254
  2. Accuracy and calibration of commercial oligonucleotide and custom cDNA microarrays.
    Nucleic Acids Res. 2002 May 15;30(10):e48 PMID: 12000853
  3. Stromal effects on mammary gland development and breast cancer.
    Science. 2002 May 10;296(5570):1046-9 PMID: 12004111
  4. Common adult stem cells in the human breast give rise to glandular and myoepithelial cell lineages: a new cell biological concept.
    Lab Invest. 2002 Jun;82(6):737-46 PMID: 12065684
  5. WISP3 is a novel tumor suppressor gene of inflammatory breast cancer.
    Oncogene. 2002 May 9;21(20):3172-80 PMID: 12082632
  6. The essential roles of the chemokine SDF-1 and its receptor CXCR4 in human stem cell homing and repopulation of transplanted immune-deficient NOD/SCID and NOD/SCID/B2m(null) mice.
    Leukemia. 2002 Oct;16(10):1992-2003 PMID: 12357350
  7. "Stemness": transcriptional profiling of embryonic and adult stem cells.
    Science. 2002 Oct 18;298(5593):597-600 PMID: 12228720
  8. A stem cell molecular signature.
    Science. 2002 Oct 18;298(5593):601-4 PMID: 12228721
  9. Functional and molecular characterisation of mammary side population cells.
    Breast Cancer Res. 2003;5(1):R1-8 PMID: 12559051
  10. Prospective identification of tumorigenic breast cancer cells.
    Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):3983-8 PMID: 12629218
  11. The influence of mammogenic hormones on serially transplanted mouse mammary gland.
    Exp Gerontol. 1971 Feb 1;6(1):95-101 PMID: 5572743
  12. Collagenous substrata regulate the nature and distribution of glycosaminoglycans produced by differentiated cultures of mouse mammary epithelial cells.
    Exp Cell Res. 1985 Feb;156(2):487-99 PMID: 3917927
  13. Interaction of mouse mammary epithelial cells with collagen substrata: regulation of casein gene expression and secretion.
    Proc Natl Acad Sci U S A. 1985 Mar;82(5):1419-23 PMID: 3856271
  14. A simplified method for passage and long-term growth of human mammary epithelial cells.
    In Vitro Cell Dev Biol. 1986 Jan;22(1):6-12 PMID: 2418007
  15. 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 Oct 1;89(19):9064-8 PMID: 1384042
  16. Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell.
    Dev Biol. 1996 Apr 10;175(1):1-13 PMID: 8608856
  17. Is there a neural stem cell in the mammalian forebrain?
    Trends Neurosci. 1996 Sep;19(9):387-93 PMID: 8873356
  18. Regulatory mechanisms in stem cell biology.
    Cell. 1997 Feb 7;88(3):287-98 PMID: 9039255
  19. An entire functional mammary gland may comprise the progeny from a single cell.
    Development. 1998 May;125(10):1921-30 PMID: 9550724
  20. Notch4 and Wnt-1 proteins function to regulate branching morphogenesis of mammary epithelial cells in an opposing fashion.
    Dev Biol. 1998 Apr 15;196(2):204-17 PMID: 9576833
  21. Expression of leukemia inhibitory factor and its receptor in breast cancer: a potential autocrine and paracrine growth regulatory mechanism.
    Breast Cancer Res Treat. 1998 Mar;48(2):165-74 PMID: 9596488
  22. Phenotypic and functional characterization in vitro of a multipotent epithelial cell present in the normal adult human breast.
    Differentiation. 1998 Aug;63(4):201-13 PMID: 9745711
  23. Mammaglobin expression in primary, metastatic, and occult breast cancer.
    Cancer Res. 1999 Jul 1;59(13):3028-31 PMID: 10397237
  24. Marrow-mindedness: a perspective on neuropoiesis.
    Trends Neurosci. 1999 Aug;22(8):348-57 PMID: 10407420
  25. Functional culture models to study mechanisms governing apoptosis in normal and malignant mammary epithelial cells.
    J Mammary Gland Biol Neoplasia. 1999 Apr;4(2):193-201 PMID: 10426398
  26. Division of labor among the alpha6beta4 integrin, beta1 integrins, and an E3 laminin receptor to signal morphogenesis and beta-casein expression in mammary epithelial cells.
    Mol Biol Cell. 1999 Sep;10(9):2817-28 PMID: 10473629
  27. Stem cells: units of development, units of regeneration, and units in evolution.
    Cell. 2000 Jan 7;100(1):157-68 PMID: 10647940
  28. Evidence that a cell-type-specific efflux pump regulates cell differentiation in Dictyostelium.
    Dev Biol. 2000 Apr 1;220(1):53-61 PMID: 10720430
  29. The tenascin family of ECM glycoproteins: structure, function, and regulation during embryonic development and tissue remodeling.
    Dev Dyn. 2000 Jun;218(2):235-59 PMID: 10842355
  30. Three to four-year-old nonpassaged EGF-responsive neural progenitor cells: proliferation, apoptosis, and DNA repair.
    Exp Neurol. 2000 Jul;164(1):200-8 PMID: 10877930
  31. Leukemia inhibitory factor (LIF) concentration modulates embryonic stem cell self-renewal and differentiation independently of proliferation.
    Biotechnol Bioeng. 2000 Sep 20;69(6):607-17 PMID: 10918135
  32. Hoechst dye efflux reveals a novel CD7(+)CD34(-) lymphoid progenitor in human umbilical cord blood.
    Blood. 2000 Sep 15;96(6):2125-33 PMID: 10979957
  33. Stem cell characteristics of transplanted rat mammary clonogens.
    Exp Cell Res. 2000 Oct 10;260(1):146-59 PMID: 11010819
  34. Normal human mammary epithelial cells spontaneously escape senescence and acquire genomic changes.
    Nature. 2001 Feb 1;409(6820):633-7 PMID: 11214324
  35. Mammary epithelial stem cells.
    Microsc Res Tech. 2001 Jan 15;52(2):190-203 PMID: 11169867
  36. Involvement of chemokine receptors in breast cancer metastasis.
    Nature. 2001 Mar 1;410(6824):50-6 PMID: 11242036
  37. A genetic analysis of neural progenitor differentiation.
    Neuron. 2001 Feb;29(2):325-39 PMID: 11239426
  38. The Hedgehog and Wnt signalling pathways in cancer.
    Nature. 2001 May 17;411(6835):349-54 PMID: 11357142
  39. From hematopoiesis to neuropoiesis: evidence of overlapping genetic programs.
    Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7934-9 PMID: 11438738
  40. The evolving concept of a stem cell: entity or function?
    Cell. 2001 Jun 29;105(7):829-41 PMID: 11439179
  41. Characterization of bipotent mammary epithelial progenitor cells in normal adult human breast tissue.
    Breast Cancer Res Treat. 2001 May;67(2):93-109 PMID: 11519870
  42. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype.
    Nat Med. 2001 Sep;7(9):1028-34 PMID: 11533706
  43. A possible role for CXCR4 and its ligand, the CXC chemokine stromal cell-derived factor-1, in the development of bone marrow metastases in neuroblastoma.
    J Immunol. 2001 Oct 15;167(8):4747-57 PMID: 11591806
  44. The interplay of matrix metalloproteinases, morphogens and growth factors is necessary for branching of mammary epithelial cells.
    Development. 2001 Aug;128(16):3117-31 PMID: 11688561
  45. Stem cells, cancer, and cancer stem cells.
    Nature. 2001 Nov 1;414(6859):105-11 PMID: 11689955
  46. MCF-10A-NeoST: a new cell system for studying cell-ECM and cell-cell interactions in breast cancer.
    Clin Cancer Res. 2001 Nov;7(11):3640-8 PMID: 11705887
  47. Signaling pathways in mammary gland development.
    Dev Cell. 2001 Oct;1(4):467-75 PMID: 11703938
  48. Wnt signaling in breast cancer: have we come full circle?
    Breast Cancer Res. 2001;3(6):351-5 PMID: 11737884
  49. Proteoglycans of the extracellular matrix and growth control.
    J Cell Physiol. 2001 Dec;189(3):266-74 PMID: 11748584
  50. The ABCG2 transporter is an efficient Hoechst 33342 efflux pump and is preferentially expressed by immature human hematopoietic progenitors.
    Blood. 2002 Jan 15;99(2):507-12 PMID: 11781231
  51. ABC transporters as phenotypic markers and functional regulators of stem cells.
    Stem Cells. 2002;20(1):11-20 PMID: 11796918
  52. Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties.
    Genes Dev. 2002 Mar 15;16(6):693-706 PMID: 11914275
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1061803
Email
gdontu@umich.edu
Published
2003-05-15
Pages
1253-70
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC196056
Subset
IM
Grants
NCI NIH HHS · P01 CA075136 · United States
NCI NIH HHS · 5P30CA46592 · United States
NCI NIH HHS · CA075136 · United States
External Links
PubMed source DOI: 10.1101/gad.1061803 PMC Full Text View journal details
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