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Side Population

  • Fariba BehbodEmail author
  • Maria d. M. Vivanco
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1293)

Abstract

The side population (SP) assay has been utilized as a method for isolation and characterization of normal and cancer stem cells from a variety of tissues. However, the SP phenotype may not be a common property of all stem cells. This chapter reviews the principle and potential pitfalls of the SP assay with an emphasis on mammary gland SP cell analysis.

Key words

Side Population (SP) Fluorescence-activated cell sorter (FACS) Stem cells Hoechst 33342 Breast cancer resistance protein (BCRP1/ABCG2) 

Notes

Acknowledgement

The authors thank K99/R00 Pathway to Independence Award: 5R00CA127462-06 (F Behbod) and the Institute of Health Carlos III (PI11/02251), the Departments of Education (PI2009-7) and Health (2010-111060), and the Department of Industry, Tourism and Trade (Etortek) of the Government of the Autonomous Community of the Basque Country (MdM Vivanco) for financial support.

References

  1. 1.
    Goodell MA et al (1996) Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. J Exp Med 183(4):1797–1806PubMedCrossRefGoogle Scholar
  2. 2.
    Zhou S et al (2001) 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 7(9):1028–1034PubMedCrossRefGoogle Scholar
  3. 3.
    Alvi AJ et al (2003) Functional and molecular characterisation of mammary side population cells. Breast Cancer Res 5(1):R1–R8PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Clayton H, Titley I, Vivanco M (2004) Growth and differentiation of progenitor/stem cells derived from the human mammary gland. Exp Cell Res 297(2):444–460PubMedCrossRefGoogle Scholar
  5. 5.
    Golebiewska A et al (2011) Critical appraisal of the side population assay in stem cell and cancer stem cell research. Cell Stem Cell 8(2):136–147PubMedCrossRefGoogle Scholar
  6. 6.
    Jonker JW et al (2005) Contribution of the ABC transporters Bcrp1 and Mdr1a/1b to the side population phenotype in mammary gland and bone marrow of mice. Stem Cells 23(8):1059–1065PubMedCrossRefGoogle Scholar
  7. 7.
    Kubota H, Avarbock MR, Brinster RL (2003) Spermatogonial stem cells share some, but not all, phenotypic and functional characteristics with other stem cells. Proc Natl Acad Sci U S A 100(11):6487–6492PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Triel C et al (2004) Side population cells in human and mouse epidermis lack stem cell characteristics. Exp Cell Res 295(1):79–90PubMedCrossRefGoogle Scholar
  9. 9.
    Iwatani H et al (2004) Hematopoietic and nonhematopoietic potentials of Hoechst(low)/side population cells isolated from adult rat kidney. Kidney Int 65(5):1604–1614PubMedCrossRefGoogle Scholar
  10. 10.
    Jonker JW et al (2005) The breast cancer resistance protein BCRP (ABCG2) concentrates drugs and carcinogenic xenotoxins into milk. Nat Med 11(2):127–129PubMedCrossRefGoogle Scholar
  11. 11.
    Lapidot T et al (1994) A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature 367(6464):645–648PubMedCrossRefGoogle Scholar
  12. 12.
    Wouters J et al (2013) The human melanoma side population displays molecular and functional characteristics of enriched chemoresistance and tumorigenesis. PLoS One 8(10):e76550PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Britton KM et al (2012) Breast cancer, side population cells and ABCG2 expression. Cancer Lett 323(1):97–105PubMedCrossRefGoogle Scholar
  14. 14.
    Mukhopadhyay P et al (2013) Heterogeneity of functional properties of clone 66 murine breast cancer cells expressing various stem cell phenotypes. PLoS One 8(11):e78725PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Natarajan K et al (2012) Role of breast cancer resistance protein (BCRP/ABCG2) in cancer drug resistance. Biochem Pharmacol 83(8):1084–1103PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Malfuson JV et al (2014) SP/drug efflux functionality of hematopoietic progenitors is controlled by mesenchymal niche through VLA-4/CD44 axis. Leukemia 28(4):853–864PubMedCrossRefGoogle Scholar
  17. 17.
    Kim JB et al (2014) Side population in LX2 cells decreased by transforming growth factor-beta. Hepatol Res 44(2):229–237PubMedCrossRefGoogle Scholar
  18. 18.
    Liu PP et al (2014) Metabolic regulation of cancer cell side population by glucose through activation of the Akt pathway. Cell Death Differ 21(1):124–135PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    To KK et al (2009) Escape from hsa-miR-519c enables drug-resistant cells to maintain high expression of ABCG2. Mol Cancer Ther 8(10):2959–2968PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Wang F et al (2010) hsa-miR-520h downregulates ABCG2 in pancreatic cancer cells to inhibit migration, invasion, and side populations. Br J Cancer 103(4):567–574PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Pan YZ, Morris ME, Yu AM (2009) MicroRNA-328 negatively regulates the expression of breast cancer resistance protein (BCRP/ABCG2) in human cancer cells. Mol Pharmacol 75(6):1374–1379PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Smalley MJ et al (2012) Isolation of mouse mammary epithelial subpopulations: a comparison of leading methods. J Mammary Gland Biol Neoplasia 17(2):91–97PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media LLC New York 2015

Authors and Affiliations

  1. 1.Pathology and Laboratory MedicineUniversity of Kansas Medical CenterKansas CityUSA
  2. 2.CIC bioGUNETechnological Park of BizkaiaDerioSpain

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