Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels


Cancer recurrence can arise owing to rare circulating cancer stem cells (CSCs) that are resistant to chemotherapies and radiotherapies. Here, we show that a double-network hydrogel can rapidly reprogramme differentiated cancer cells into CSCs. Spheroids expressing elevated levels of the stemness genes Sox2, Oct3/4 and Nanog formed within 24 h of seeding the gel with cells from any of six human cancer cell lines or with brain cancer cells resected from patients with glioblastoma. Human brain cancer cells cultured on the double-network hydrogel and intracranially injected in immunodeficient mice led to higher tumorigenicity than brain cancer cells cultured on single-network gels. We also show that the double-network gel induced the phosphorylation of tyrosine kinases, that gel-induced CSCs from primary brain cancer cells were eradicated by an inhibitor of the platelet-derived growth factor receptor, and that calcium channel receptors and the protein osteopontin were essential for the regulation of gel-mediated induction of stemness in brain cancer cells.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: DN gels rapidly induced CSCs of six human cancer cell lines.
Fig. 2: DN gels rapidly induced CSCs of human brain cancer cell lines and primary culture cells.
Fig. 3: Analysis of hydrogels for induction of CSCs and comparison of expression profiles of CSCs induced by DN gel and neurosphere culture conditions.
Fig. 4: Tumorigenic potential of DN gel-induced sphere-forming brain cancer cells in vivo.
Fig. 5: Analysis of DN gel-activated intracellular signalling pathways.
Fig. 6: Requirement for OPN for induction of brain CSCs by DN gels.
Fig. 7: DN gels induced PDGFR-expressing CSCs, which were eradicated by PDGFR inhibitor.
Fig. 8: Molecular mechanisms of the hydrogel-activated reprogramming phenomenon.

Data availability

The main data supporting the results of this study are available within the paper and its Supplementary Information. All raw data used to make the graphs are available from Figshare with the identifier


  1. 1.

    Torre, L. A. et al. Global cancer statistics, 2012. CA Cancer J. Clin. 65, 87–108 (2015).

    Article  Google Scholar 

  2. 2.

    Siegel, R. L., Miller, K. D. & Jemal, A. Cancer statistics, 2017. CA Cancer J. Clin. 67, 7–30 (2017).

    Article  Google Scholar 

  3. 3.

    Kreso, A. & Dick, J. E. Evolution of the cancer stem cell model. Cell Stem Cell 14, 275–291 (2014).

    CAS  Article  Google Scholar 

  4. 4.

    Reya, T., Morrison, S. J., Clarke, M. F. & Weissman, I. L. Stem cells, cancer, and cancer stem cells. Nature 414, 105–111 (2001).

    CAS  Article  Google Scholar 

  5. 5.

    Batlle, E. & Clevers, H. Cancer stem cells revisited. Nat. Med. 23, 1124–1134 (2017).

    CAS  Article  Google Scholar 

  6. 6.

    Clevers, H. The cancer stem cell: premises, promises and challenges. Nat. Med. 17, 313–319 (2011).

    CAS  Article  Google Scholar 

  7. 7.

    Boiko, A. D. et al. Human melanoma-initiating cells express neural crest nerve growth factor receptor CD271. Nature 466, 133–137 (2010).

    CAS  Article  Google Scholar 

  8. 8.

    Stuckey, D. W. & Shah, K. Stem cell-based therapies for cancer treatment: separating hope from hype. Nat. Rev. Cancer 14, 683–691 (2014).

    CAS  Article  Google Scholar 

  9. 9.

    Madl, C. M., Heilshorn, S. C. & Blau, H. M. Bioengineering strategies to accelerate stem cell therapeutics. Nature 557, 335–342 (2018).

    CAS  Article  Google Scholar 

  10. 10.

    Vining, K. H. & Mooney, D. J. Mechanical forces direct stem cell behaviour in development and regeneration. Nat. Rev. Mol. Cell Biol. 18, 728–742 (2017).

    CAS  Article  Google Scholar 

  11. 11.

    Gong, J. P. Materials science. Materials both tough and soft. Science 344, 161–162 (2014).

    CAS  Article  Google Scholar 

  12. 12.

    Goto, K. et al. Synthetic PAMPS gel activates BMP/Smad signaling pathway in ATDC5 cells, which plays a significant role in the gel-induced chondrogenic differentiation. J. Biomed. Mater. Res. A 104, 734–746 (2015).

    Article  Google Scholar 

  13. 13.

    Yasuda, K. et al. A novel double-network hydrogel induces spontaneous articular cartilage regeneration in vivo in a large osteochondral defect. Macromol. Biosci. 9, 307–316 (2009).

    CAS  Article  Google Scholar 

  14. 14.

    Imabuchi, R. et al. Gene expression profile of the cartilage tissue spontaneously regenerated in vivo by using a novel double-network gel: comparisons with the normal articular cartilage. BMC Musculoskelet. Disord. 12, 213 (2011).

    CAS  Article  Google Scholar 

  15. 15.

    Bendall, S. C. et al. IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitro. Nature 448, 1015–1021 (2007).

    CAS  Article  Google Scholar 

  16. 16.

    Jabbari, E., Sarvestani, S. K., Daneshian, L. & Moeinzadeh, S. Optimum 3D matrix stiffness for maintenance of cancer stem cells is dependent on tissue origin of cancer cells. PLoS ONE 10, e0132377 (2015).

    Article  Google Scholar 

  17. 17.

    Tabu, K. et al. A synthetic polymer scaffold reveals the self-maintenance strategies of rat glioma stem cells by organization of the advantageous niche. Stem Cells 34, 1151–1162 (2016).

    CAS  Article  Google Scholar 

  18. 18.

    Chen, J., McKay, R. M. & Parada, L. F. Malignant glioma: lessons from genomics, mouse models, and stem cells. Cell 149, 36–47 (2012).

    CAS  Article  Google Scholar 

  19. 19.

    Vescovi, A. L., Galli, R. & Reynolds, B. A. Brain tumour stem cells. Nat. Rev. Cancer 6, 425–436 (2006).

    CAS  Article  Google Scholar 

  20. 20.

    Lerner, R. G. et al. Targeting a Plk1-controlled polarity checkpoint in therapy-resistant glioblastoma-propagating cells. Cancer Res. 75, 5355–5366 (2015).

    CAS  Article  Google Scholar 

  21. 21.

    Li, J. et al. KPNA2 promotes metabolic reprogramming in glioblastomas by regulation of c-myc. J. Exp. Clin. Cancer Res. 37, 194 (2018).

    Article  Google Scholar 

  22. 22.

    Frauenlob, M. et al. Modulation and characterization of the double network hydrogel surface-bulk transition. Macromolecules 52, 6704–6713 (2019).

    CAS  Article  Google Scholar 

  23. 23.

    Boccaccio, C. & Comoglio, P. M. The MET oncogene in glioblastoma stem cells: implications as a diagnostic marker and a therapeutic target. Cancer Res. 73, 3193–3199 (2013).

    CAS  Article  Google Scholar 

  24. 24.

    Kohsaka, S. et al. Epiregulin enhances tumorigenicity by activating the ERK/MAPK pathway in glioblastoma. Neuro-Oncol. 16, 960–970 (2014).

    CAS  Article  Google Scholar 

  25. 25.

    Pietras, A. et al. Osteopontin-CD44 signaling in the glioma perivascular niche enhances cancer stem cell phenotypes and promotes aggressive tumor growth. Cell Stem Cell 14, 357–369 (2014).

    CAS  Article  Google Scholar 

  26. 26.

    Kijewska, M. et al. The embryonic type of SPP1 transcriptional regulation is re-activated in glioblastoma. Oncotarget 8, 16340–16355 (2017).

    Article  Google Scholar 

  27. 27.

    Elbediwy, A., Vincent-Mistiaen, Z. I. & Thompson, B. J. YAP and TAZ in epithelial stem cells: a sensor for cell polarity, mechanical forces and tissue damage. Bioessays 38, 644–653 (2016).

    CAS  Article  Google Scholar 

  28. 28.

    Halder, G., Dupont, S. & Piccolo, S. Transduction of mechanical and cytoskeletal cues by YAP and TAZ. Nat. Rev. Mol. Cell Biol. 13, 591–600 (2012).

    CAS  Article  Google Scholar 

  29. 29.

    Song, J. et al. β1 integrin mediates colorectal cancer cell proliferation and migration through regulation of the Hedgehog pathway. Tumour Biol. 36, 2013–2021 (2015).

    CAS  Article  Google Scholar 

  30. 30.

    Gong, J. P., Katsuyama, Y., Kurokawa, T. & Osada, Y. Double-network hydrogels with extremely high mechanical strength. Adv. Mater. 15, 1155–1158 (2003).

    CAS  Article  Google Scholar 

  31. 31.

    Momozaki, N. et al. Suppression of anchorage-independent growth of human glioblastoma cell by major histocompatibility complex class I gene-transfection. J. Neurosurg. 76, 845–849 (1992).

    CAS  Article  Google Scholar 

  32. 32.

    Kawai, A. et al. Establishment and characterization of a biphasic synovial sarcoma cell line, SYO-1. Cancer Lett. 204, 105–113 (2004).

    CAS  Article  Google Scholar 

  33. 33.

    Nojima, T. et al. Morphological and cytogenetic studies of a human synovial sarcoma xenotransplanted into nude mice. Acta Pathol. Jpn 40, 486–493 (1990).

    CAS  PubMed  Google Scholar 

  34. 34.

    Liang, S. et al. Isolation and characterization of human breast cancer cells with SOX2 promoter activity. Biochem. Biophys. Res. Commun. 437, 205–211 (2013).

    CAS  Article  Google Scholar 

  35. 35.

    Mizuno, H., Kitada, K., Nakai, K. & Sarai, A. PrognoScan: a new database for meta-analysis of the prognostic value of genes. BMC Med. Genomics 2, 18 (2009).

    Article  Google Scholar 

Download references


We thank K. Hida (Hokkaido University, Japan) for the pCSII-CMV-tdTomato-Luc2 plasmid, S. Kon (Fukuyama University, Japan) for the anti-OPN antibody (clone 35B6), M. Sudol (National University of Singapore) for p2×FLAG-CMV2-YAP1, K. Tabuchi (Saga University, Japan) for the GBM cell line KMG4, A. Kawai (National Cancer Center, Japan) for the synovial sarcoma cell line SYO-1, A. Hirota (Hokkaido University, Japan) for the human induced pluripotent stem cells and Y. Hane (Hokkaido University) for gel synthesis. We also thank the medical students R. Nabeshima, K. Aoyama, T. Kurai and T. Ishizuka (Hokkaido University) for useful discussions. This work is supported by the Global Station for Soft Matter (a project of the Global Institution for Collaborative Research and Education at Hokkaido University) and, in part, by grants from MEXT (19H01171 to S.T., and 15K15106 and 18K07059 to M.T.), and AMED (20cm0106571h0001 and 21cm0106571h0002 to S.T.). The Institute for Chemical Reaction Design and Discovery (ICReDD) was established by the World Premier International Research Center Initiative (WPI), MEXT, Japan. This study used IVIS with support from the Global Center for Biomedical Science and Engineering, Faculty of Medicine, Hokkaido University.

Author information




J.S. performed all of the in vitro experiments with supervision from M.T. and S.S. L.W. performed the animal experiments. S.A. and Y.O. performed the microarray analysis. H.S. performed the calcium ion-channel analysis. T.K. and M.F. generated the hydrogels. K.Y. and J.P.G. organized the projects related to hydrogels at Hokkaido University. S. Kohsaka, S.Kojima, T.U. and H.M. performed the single-cell whole-transcriptome analysis. K.K. performed the immunofluorescence analysis. S.T. designed the entire study and wrote the manuscript.

Corresponding author

Correspondence to Shinya Tanaka.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary methods, figures, tables and video captions.

Reporting Summary

Supplementary Video 1

Rapid induction of spheroid formation on a DN gel.

Supplementary Video 2

SOX2-expressing cells in a spheroid formed on a DN gel.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Suzuka, J., Tsuda, M., Wang, L. et al. Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels. Nat Biomed Eng (2021).

Download citation

Further reading


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing