Cancer Stem Cells – Basic Biological Properties and Experimental Approaches


Cancer stem cells (CSCs) are defined as a self‐renewing and self‐protecting subpopulation of tumour cells that can differentiate into all other tumour cell types found within a tumour. They can be prospectively identified and purified, either on the basis of specific cell surface marker expression, or by virtue of their biological properties; their self‐protection has been harnessed to develop several purification strategies, for example, side population or Aldefluor assays. Other prominent biological properties of CSCs include their exclusive clonogenicity and tumourigenicity. Many of these basic biological characteristics are shared between normal and CSCs, as are multiple signalling pathways regulating self‐renewal and differentiation. Unlike normal stem cells, CSCs often seem to be distinctly less stable, dynamically modulating their stemness and differentiation according to various environmental signals and/or genetic and epigenetic changes accumulated during the carcinogenic process. This plasticity of CSCs might underlie the prominent biological characteristics of malignant tumours, especially their propensity to metastasise.

Key Concepts:

  • Stem cells are long‐lived and protect their genome.

  • Existence of long‐lived, undifferentiated and self‐protecting cells could be indirectly inferred in tumours.

  • Cancer stem cells express specific cell surface markers and are amenable to differential staining procedures.

  • Cancer stem cells frequently exploit similar signalling pathways as embryonic stem cells and normal adult stem cells.

  • Hypoxia activates a multifaceted stem cell programme in tumours.

  • Cancer stem cell phenotype can be unstable, with repeated or even ongoing differentiation and dedifferentiation.

Keywords: cancer stem cells; cancer stem cell markers; stem cell self‐renewal and differentiation; cancer stem cell plasticity

Figure 1.

Two text‐book models of cancer cell growth and heterogeneity. The stochastic model (a) assumes that different subpopulations of tumour cells continuously arise and disappear by evolutionary processes like clonal selection. It is unable to prospectively isolate any subpopulation of tumour cells uniquely responsible for both perpetuating tumour growth and its spread to a secondary site, whereas in the hierarchical tumour model (b) only the CSCs are able to both perpetuate and transfer the tumour, by virtue of their exclusive self‐renewing ability.

Figure 2.

Hypoxia can impact on stem cell biology at multiple different levels, both by participating in the induction of Notch‐regulated genes (the leftmost pathway) and by directly inducing multiple genes involved in various aspects of stemness. Note that the individual effects can be partly contradictory – hypoxia can thus both induce the epithelial‐mesenchymal transition (EMT) and strenghten the epithelial character of cells.



Al‐Hajj M , Wicha MS , Benito‐Hernandez A , Morrison SJ and Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proceedings of the National Academy of Sciences of the USA 100: 3983–3988.

Barker N , van Es JH , Kuipers J et al. (2007) Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449: 1003–1007.

Bedogni B , Warneke JA , Nickoloff BJ , Giaccia AJ and Powell MB (2008) Notch1 is an effector of Akt and hypoxia in melanoma development. Journal of Clinical Investigation 118: 3660–3670.

Bhattacharyya S , Kumar A and Lal Khanduja K (2012) The voyage of stem cell toward terminal differentiation: a brief overview. Acta Biochimica et Biophysica Sinica 44: 463–475.

Biddle A and Mackenzie IC (2012) Cancer stem cells and EMT in carcinoma. Cancer and Metastasis Reviews 31: 285–293.

Bonizzi G , Cicalese A , Insinga A and Pelicci PG (2012) The emerging role of p53 in stem cells. Trends in Molecular Medicine 18: 6–12.

Bourguignon LY , Wong G , Earle C and Chen L (2012) Hyaluronan‐CD44v3 interaction with Oct4‐Sox2‐Nanog promotes miR‐302 expression leading to self‐renewal, clonal formation, and cisplatin resistance in cancer stem cells from head and neck squamous cell carcinoma. Journal of Biological Chemistry 287: 32800–32824.

Brabletz T , Jung A , Spaderna S , Hlubek F and Kirchner T (2005) Opinion: migrating cancer stem cells – an integrated concept of malignant tumour progression. Nature Reviews Cancer 5: 744–749.

Buchstaller J , Quintana E and Morrison SJ (2008) Cancer stem cells. In: Mendelson J , Howley PM , Israel MA , Gray JW and Thompson CB (eds) The Molecular Basis of Cancer, pp 141–154. Philadelphia: Saunders Elsevier.

Campos LS (2004) Neurospheres: insights into neural stem cell biology. Journal of Neuroscience Research 78: 761–769.

Celià‐Terrassa T , Meca‐Cortés O , Mateo F et al. (2012) Epithelial‐mesenchymal transition can suppress major attributes of human epithelial tumor‐initiating cells. Journal of Clinical Investigation 122: 1849–1868.

Chan KS , Espinosa I , Chao M et al. (2009) Identification, molecular characterization, clinical prognosis, and therapeutic targeting of human bladder tumor-initiating cells. Proceedings of the National Academy of Sciences of the USA 106: 14016–14021.

Christgen M , Geffers R , Ballmaier M et al. (2010) Down‐regulation of the fetal stem cell factor SOX17 by H33342: a mechanism responsible for differential gene expression in breast cancer side population cells. Journal of Biological Chemistry 285: 6412–6418.

Clarkson BD (1969) Review of recent studies of cellular proliferation in acute leukemia. Journal of the National Cancer Institute Monographs 30: 81–120.

Clevers H and Nusse R (2012) Wnt/β‐catenin signaling and disease. Cell 149: 1192–1205.

Desmarais JA , Hoffmann MJ , Bingham G et al. (2012) Human embryonic stem cells fail to activate CHK1 and commit to apoptosis in response to DNA replication stress. Stem Cells 30: 1385–1393.

Godar S , Ince TA , Bell GW et al. (2008) Growth‐inhibitory and tumor‐ suppressive functions of p53 depend on its repression of CD44 expression. Cell 134: 62–73.

Greve B , Kelsch R , Spaniol K , Eich HT and Götte M (2012) Flow cytometry in cancer stem cell analysis and separation. Cytometry A 81: 284–293.

Grimshaw MJ , Cooper L , Papazisis K et al. (2008) Mammosphere culture of metastatic breast cancer cells enriches for tumorigenic breast cancer cells. Breast Cancer Research 10: R52.

Grosse‐Gehling P , Fargeas CA , Dittfeld C et al. (2013) CD133 as a biomarker for putative cancer stem cells in solid tumours: limitations, problems and challenges. Journal of Pathology 229: 355–378.

Hatina J (2012) The dynamics of cancer stem cells. Neoplasma 59: 700–707.

Hatina J , Schulz WA , Fischer J et al. (2007) Tumour stem cells – a new concept in tumour biology [In German] Deutsche Medizinische Wochenschrift 132: 1629–1632.

Hatina J and Schulz WA (2012) Stem cells in the biology of normal urothelium and urothelial carcinoma. Neoplasma 59: 728–736.

Hermann PC , Huber SL , Herrler T et al. (2007) Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 1: 313–323.

John S , Stuelten CH and Niederhuber JE (2009) Frontiers of Cancer Research: The Metastasis. In: Leong SPL (ed.) From Local Invasion to Metastatic Cancer. Current Clinical Oncology, pp 3–10. New York: Humana Press.

Jones PH (2010) Stem cell fate in proliferating tissues: equal odds in a game of chance. Developmental Cell 19: 489–490.

Kajstura J , Bai Y , Cappetta D et al. (2012) Tracking chromatid segregation to identify human cardiac stem cells that regenerate extensively the infarcted myocardium. Circulation Research 111: 894–906.

Kashyap V , Rezende NC , Scotland KB et al. (2009) Regulation of stem cell pluripotency and differentiation involves a mutual regulatory circuit of the NANOG, OCT4, and SOX2 pluripotency transcription factors with polycomb repressive complexes and stem cell microRNAs. Stem Cells and Development 18: 1093–1108.

Kim JB , Ko E , Han W et al. (2008) CD24 cross‐linking induces apoptosis in, and inhibits migration of, MCF‐7 breast cancer cells. BMC Cancer 8: 118.

Kim MY , Oskarsson T , Acharyya S et al. (2009) Tumor self‐seeding by circulating cancer cells. Cell 139: 1315–1326.

Lapidot T , Sirard C , Vormoor J et al. (1994) A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature 367: 645–648.

Li H , Chen X , Calhoun‐Davis T , Claypool K and Tang DG (2008) PC3 human prostate carcinoma cell holoclones contain self‐renewing tumor‐initiating cells. Cancer Research 68: 1820–1825.

Liu C , Kelnar K , Liu B et al. (2011) The microRNA miR‐34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. Nature Medicine 17: 211–215.

Liu Q , Yu F-Y , Tang W , Su S-C and Song E-W (2012) Cancer stem cell. In: Liu X-Y , Pestka S and Shi Y-F (eds) Recent Advances in Cancer Research and Therapy, pp 173–196. Beijing, China: Tsinghua University Press and Elsevier.

Locke M , Heywood M , Fawell S and Mackenzie IC (2005) Retention of intrinsic stem cell hierarchies in carcinoma‐derived cell lines. Cancer Research 65: 8944–8950.

Ma J , Meng Y , Kwiatkowski DJ et al. (2010) Mammalian target of rapamycin regulates murine and human cell differentiation through STAT3/p63/Jagged/Notch cascade. Journal of Clinical Investigation 120: 103–114.

Mani SA , Guo W , Liao MJ et al. (2008) The epithelial‐mesenchymal transition generates cells with properties of stem cells. Cell 133: 704–715.

Marotta LL , Almendro V , Marusyk A et al. (2011) The JAK2/STAT3 signaling pathway is required for growth of CD44+CD24− stem cell‐like breast cancer cells in human tumors. Journal of Clinical Investigation 121: 2723–2735.

Mohyeldin A , Garzón‐Muvdi T and Quiñones‐Hinojosa A (2010) Oxygen in stem cell biology: a critical component of the stem cell niche. Cell Stem Cell 7: 150–161.

Nguyen LV , Vanner R , Dirks P and Eaves CJ (2012) Cancer stem cells: an evolving concept. Nature Reviews Cancer 12: 133–143.

Park JT , Chen X , Tropè CG et al. (2010) Notch3 overexpression is related to the recurrence of ovarian cancer and confers resistance to carboplatin. American Journal of Pathology 177: 1087–1094.

Proweller A , Tu L , Lepore JJ et al. (2006) Impaired notch signaling promotes de novo squamous cell carcinoma formation. Cancer Research 66: 7438–7444.

Quail DF , Taylor MJ and Postovit LM (2012) Microenvironmental regulation of cancer stem cell phenotypes. Current Stem Cell Research & Therapy 7: 197–216.

Quintana E , Shackleton M , Sabel MS et al. (2008) Efficient tumour formation by single human melanoma cells. Nature 456: 593–598.

Quyn AJ , Appleton PL , Carey FA et al. (2010) Spindle orientation bias in gut epithelial stem cell compartments is lost in precancerous tissue. Cell Stem Cell 6: 175–181.

Ramos A and Camargo FD (2012) The Hippo signaling pathway and stem cell biology. Trends in Cell Biology 22: 339–346.

Rando TA (2007) The immortal strand hypothesis: segregation and reconstruction. Cell 129: 1239–1243.

Ricci‐Vitiani L , Lombardi DG , Pilozzi E et al. (2007) Identification and expansion of human colon‐cancer‐initiating cells. Nature 445: 111–115.

Sainson RC and Harris AL (2006) Hypoxia‐regulated differentiation: let's step it up a Notch. Trends in Molecular Medicine 12: 141–143.

Sell S (2009) History of cancer stem cells. In: Rajasekhar VK and Vemuri MC (eds) Regulatory Networks in Stem Cells. Stem Cell Biology and Regenerative Medicine, pp 495–503. New York: Humana Press.

Shackleton M , Quintana E , Fearon ER and Morrison SJ (2009) Heterogeneity in cancer: cancer stem cells versus clonal evolution. Cell 138: 822–829.

She JJ , Zhang PG , Wang ZM , Gan WM and Che XM (2008) Identification of side population cells from bladder cancer cells by DyeCycle Violet staining. Cancer Biology & Therapy 7: 1663–1668.

Shmelkov SV , Butler JM , Hooper AT et al. (2008) CD133 expression is not restricted to stem cells, and both CD133+ and CD133− metastatic colon cancer cells initiate tumors. Journal of Clinical Investigation 118: 2111–2120.

Singh SK , Clarke ID , Terasaki M et al. (2003) Identification of a cancer stem cell in human brain tumors. Cancer Research 63: 5821–5828.

South AP , Cho RJ and Aster JC (2012) The double‐edged sword of Notch signaling in cancer. Seminars in Cell & Developmental Biology 23: 458–464.

Taussig DC , Miraki‐Moud F , Anjos‐Afonso F et al. (2008) Anti‐CD38 antibody‐mediated clearance of human repopulating cells masks the heterogeneity of leukemia‐initiating cells. Blood 112: 568–575.

The Economist (2008) Cancer stem cells: the root of all evil? The Economist 388: 84–86.

Tsai JH , Donaher JL , Murphy DA , Chau S and Yang J (2012) Spatiotemporal regulation of epithelial‐mesenchymal transition is essential for squamous cell carcinoma metastasis. Cancer Cell 22: 725–736.

Vogt PK and Hart JR (2011) PI3K and STAT3: a new alliance. Cancer Discovery 1: 481–486.

Wellner U , Schubert J , Burk UC et al. (2009) The EMT‐activator ZEB1 promotes tumorigenicity by repressing stemness‐inhibiting microRNAs. Nature Cell Biology 11: 1487–1495.

Xu Y , He K and Goldkorn A (2011) Telomerase targeted therapy in cancer and cancer stem cells. Clinical Advances in Hematology & Oncology 9: 442–455.

Zeimet AG , Reimer D , Sopper S et al. (2012) Ovarian cancer stem cells. Neoplasma 59: 747–755.

Zöller M (2011) CD44: can a cancer‐initiating cell profit from an abundantly expressed molecule? Nature Reviews Cancer 11: 254–267.

Further Reading

Alison MR , Lin WR , Lim SM and Nicholson LJ (2012) Cancer stem cells: in the line of fire. Cancer Treatment Reviews 38: 589–598.

Bagley RG and Teicher BA (eds) Stem Cells and Cancer. New York: Humana Press 2009.

Christgen M , Ballmaier M , Lehmann U and Kreipe H (2012) Detection of putative cancer stem cells of the side population phenotype in human tumor cell cultures. Methods in Molecular Biology 878: 201–215.

Hjelmeland AB , Lathia JD , Sathornsumetee S and Rich JN (2011) Twisted tango: brain tumor neurovascular interactions. Nature Neuroscience 14: 1375–1381.

Karlic H , Herrmann H , Schulenburg A et al. (2010) Tumor stem cell research – basis and challenge for diagnosis and therapy [In German]. Wiener Klinische Wochenschrift 122: 423–436.

Keysar SB and Jimeno A (2010) More than markers: biological significance of cancer stem cell‐defining molecules. Molecular Cancer Therapeutics 9: 2450–2457.

Liu C and Tang DG (2011) MicroRNA regulation of cancer stem cells. Cancer Research 71: 5950–5954.

Magee JA , Piskounova E and Morrison SJ (2012) Cancer stem cells: impact, heterogeneity, and uncertainty. Cancer Cell 21: 283–296.

Rahman M , Deleyrolle L , Vedam‐Mai V et al. (2011) The cancer stem cell hypothesis: failures and pitfalls. Neurosurgery 68: 531–545.

Tirino V , Desiderio V , Paino F et al. (2013) Cancer stem cells in solid tumors: an overview and new approaches for their isolation and characterization. FASEB Journal 27: 13–24.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Hatina, Jiří, Fernandes, Maria Inès, Hoffmann, Michèle J, and Zeimet, Alain G(Sep 2013) Cancer Stem Cells – Basic Biological Properties and Experimental Approaches. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0021164.pub2]