How to Assess Drug Resistance in Cancer Stem Cells

  • Maria Laura De Angelis
  • Ruggero De Maria
  • Marta Baiocchi
Part of the Methods in Molecular Biology book series (MIMB, volume 1692)


Banks of genetically characterized cancer stem cells (CSCs) isolated from individual patients and grown as spheroids offer an invaluable approach to identify genetic determinants of drug resistance versus sensitivity, and to study new stem cell-directed therapies. Here, we describe our standardized procedure for in vitro drug screening on colorectal CSCs, taking irinotecan as an example.

Key words

CSC Cancer stem cell culture Drug screening Drug resistance In vitro assays Colorectal cancer 



This work was financially supported by the Italian Association for Cancer Research (AIRC) 5 per Mille 9979 grant to RDM.


  1. 1.
    Oskarsson T, Batlle E, Massague J (2014) Metastatic stem cells: sources, niches, and vital pathways. Cell Stem Cell 14(3):306–321. doi: 10.1016/j.stem.2014.02.002 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Zeuner A, Todaro M, Stassi G, De Maria R (2014) Colorectal cancer stem cells: from the crypt to the clinic. Cell Stem Cell 15(6):692–705. doi: 10.1016/j.stem.2014.11.012 CrossRefPubMedGoogle Scholar
  3. 3.
    Todaro M, Alea MP, Di Stefano AB, Cammareri P, Vermeulen L, Iovino F, Tripodo C, Russo A, Gulotta G, Medema JP, Stassi G (2007) Colon Cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell 1(4):389–402. doi: 10.1016/j.stem.2007.08.001 CrossRefPubMedGoogle Scholar
  4. 4.
    Cammareri P, Scopelliti A, Todaro M, Eterno V, Francescangeli F, Moyer MP, Agrusa A, Dieli F, Zeuner A, Stassi G (2010) Aurora-A is essential for the tumorigenic capacity and chemoresistance of colorectal cancer stem cells. Cancer Res 70(11):4655–4665. doi: 10.1158/0008-5472.Can-09-3953 CrossRefPubMedGoogle Scholar
  5. 5.
    Kreso A, O'Brien CA, van Galen P, Gan OI, Notta F, Brown AM, Ng K, Ma J, Wienholds E, Dunant C, Pollett A, Gallinger S, McPherson J, Mullighan CG, Shibata D, Dick JE (2013) Variable clonal repopulation dynamics influence chemotherapy response in colorectal cancer. Science 339(6119):543–548. doi: 10.1126/science.1227670 CrossRefPubMedGoogle Scholar
  6. 6.
    Colak S, Zimberlin CD, Fessler E, Hogdal L, Prasetyanti PR, Grandela CM, Letai A, Medema JP (2014) Decreased mitochondrial priming determines chemoresistance of colon cancer stem cells. Cell Death Differ 21(7):1170–1177. doi: 10.1038/cdd.2014.37 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    De Angelis ML, Zeuner A, Policicchio E, Russo G, Bruselles A, Signore M, Vitale S, De Luca G, Pilozzi E, Boe A, Stassi G, Ricci-Vitiani L, Amoreo CA, Pagliuca A, Francescangeli F, Tartaglia M, De Maria R, Baiocchi M (2016) Cancer stem cell-based models of colorectal cancer reveal molecular determinants of therapy resistance. Stem Cells Transl Med 5(4):511–523. doi: 10.5966/sctm.2015-0214 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Gupta PB, Onder TT, Jiang G, Tao K, Kuperwasser C, Weinberg RA, Lander ES (2009) Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell 138(4):645–659. doi: 10.1016/j.cell.2009.06.034 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Kreso A, van Galen P, Pedley NM, Lima-Fernandes E, Frelin C, Davis T, Cao L, Baiazitov R, Du W, Sydorenko N, Moon YC, Gibson L, Wang Y, Leung C, Iscove NN, Arrowsmith CH, Szentgyorgyi E, Gallinger S, Dick JE, O'Brien CA (2014) Self-renewal as a therapeutic target in human colorectal cancer. Nat Med 20(1):29–36. doi: 10.1038/nm.3418 CrossRefPubMedGoogle Scholar
  10. 10.
    Gao H, Korn JM, Ferretti S, Monahan JE, Wang Y, Singh M, Zhang C, Schnell C, Yang G, Zhang Y, Balbin OA, Barbe S, Cai H, Casey F, Chatterjee S, Chiang DY, Chuai S, Cogan SM, Collins SD, Dammassa E, Ebel N, Embry M, Green J, Kauffmann A, Kowal C, Leary RJ, Lehar J, Liang Y, Loo A, Lorenzana E, Robert McDonald E 3rd, ME ML, Merkin J, Meyer R, Naylor TL, Patawaran M, Reddy A, Roelli C, Ruddy DA, Salangsang F, Santacroce F, Singh AP, Tang Y, Tinetto W, Tobler S, Velazquez R, Venkatesan K, Von Arx F, Wang HQ, Wang Z, Wiesmann M, Wyss D, Xu F, Bitter H, Atadja P, Lees E, Hofmann F, Li E, Keen N, Cozens R, Jensen MR, Pryer NK, Williams JA, Sellers WR (2015) High-throughput screening using patient-derived tumor xenografts to predict clinical trial drug response. Nat Med 21(11):1318–1325. doi: 10.1038/nm.3954 CrossRefPubMedGoogle Scholar
  11. 11.
    van de Wetering M, Francies HE, Francis JM, Bounova G, Iorio F, Pronk A, van Houdt W, van Gorp J, Taylor-Weiner A, Kester L, McLaren-Douglas A, Blokker J, Jaksani S, Bartfeld S, Volckman R, van Sluis P, Li VSW, Seepo S, Pedamallu CS, Cibulskis K, Carter SL, McKenna A, Lawrence MS, Lichtenstein L, Stewart C, Koster J, Versteeg R, van Oudenaarden A, Saez-Rodriguez J, Vries RGJ, Getz G, Wessels L, Stratton MR, McDermott U, Meyerson M, Garnett MJ, Clevers H (2015) Prospective derivation of a living organoid biobank of colorectal cancer patients. Cell 161(4):933–945. doi: 10.1016/j.cell.2015.03.053 CrossRefPubMedGoogle Scholar
  12. 12.
    Marcucci F, Stassi G, De Maria R (2016) Epithelial-mesenchymal transition: a new target in anticancer drug discovery. Nat Rev Drug Discov 15(5):311–325. doi: 10.1038/nrd.2015.13 CrossRefPubMedGoogle Scholar
  13. 13.
    Baiocchi M, Biffoni M, Ricci-Vitiani L, Pilozzi E, De Maria R (2010) New models for cancer research: human cancer stem cell xenografts. Curr Opin Pharmacol 10(4):380–384. doi: 10.1016/j.coph.2010.05.002 CrossRefPubMedGoogle Scholar
  14. 14.
    Galli R, Gritti A, Vescovi AL (2008) Adult neural stem cells. Methods Mol Biol 438:67–84. doi: 10.1007/978-1-59745-133-8_7 CrossRefPubMedGoogle Scholar
  15. 15.
    Sato T, Stange DE, Ferrante M, Vries RGJ, van Es JH, van den Brink S, van Houdt WJ, Pronk A, van Gorp J, Siersema PD, Clevers H (2011) Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and barrett's epithelium. Gastroenterology 141(5):1762–1772. doi: 10.1053/j.gastro.2011.07.050 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Maria Laura De Angelis
    • 1
  • Ruggero De Maria
    • 2
  • Marta Baiocchi
    • 1
  1. 1.Department of Oncology and Molecular MedicineIstituto Superiore di SanitàRomeItaly
  2. 2.Department of General PathologyUniversità Cattolica del Sacro CuoreRomeItaly

Personalised recommendations