Skip to main content

Thank you for visiting nature.com. 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.

Looking to the future following 10 years of induced pluripotent stem cell technologies

Abstract

The development of induced pluripotent stem cells (iPSCs) has fundamentally changed our view on developmental cell-fate determination and led to a cascade of technological innovations in regenerative medicine. Here we provide an overview of the progress in the field over the past decade, as well as our perspective on future directions and clinical implications of iPSC technology.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Alternative reprogramming factors and facilitators of reprogramming.
Figure 2: Selected milestones in the development of iPSC technology and future perspectives.

References

  1. Gurdon, J.B., Elsdale, T.R. & Fischberg, M. Sexually mature individuals of Xenopus laevis from the transplantation of single somatic nuclei. Nature 182, 64–65 (1958).

    CAS  Article  PubMed  Google Scholar 

  2. Gurdon, J.B. The developmental capacity of nuclei taken from intestinal epithelium cells of feeding tadpoles. J. Embryol. Exp. Morphol. 10, 622–640 (1962).

    CAS  PubMed  Google Scholar 

  3. Evans, M.J. & Kaufman, M.H. Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–156 (1981).

    CAS  Article  PubMed  Google Scholar 

  4. Martin, G.R. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc. Natl. Acad. Sci. USA 78, 7634–7638 (1981).

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  5. Tada, M., Takahama, Y., Abe, K., Nakatsuji, N. & Tada, T. Nuclear reprogramming of somatic cells by in vitro hybridization with ES cells. Curr. Biol. 11, 1553–1558 (2001).

    CAS  Article  PubMed  Google Scholar 

  6. Cowan, C.A., Atienza, J., Melton, D.A. & Eggan, K. Nuclear reprogramming of somatic cells after fusion with human embryonic stem cells. Science 309, 1369–1373 (2005).

    CAS  Article  PubMed  Google Scholar 

  7. Davis, R.L., Weintraub, H. & Lassar, A.B. Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell 51, 987–1000 (1987).

    CAS  Article  PubMed  Google Scholar 

  8. Kulessa, H., Frampton, J. & Graf, T. GATA-1 reprograms avian myelomonocytic cell lines into eosinophils, thromboblasts, and erythroblasts. Genes Dev. 9, 1250–1262 (1995).

    CAS  Article  PubMed  Google Scholar 

  9. Xie, H., Ye, M., Feng, R. & Graf, T. Stepwise reprogramming of B cells into macrophages. Cell 117, 663–676 (2004).

    CAS  Article  PubMed  Google Scholar 

  10. Mitsui, K. et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631–642 (2003).

    CAS  Article  PubMed  Google Scholar 

  11. Sato, N., Meijer, L., Skaltsounis, L., Greengard, P. & Brivanlou, A.H. Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor. Nat. Med. 10, 55–63 (2004).

    CAS  Article  PubMed  Google Scholar 

  12. Cartwright, P. et al. LIF/STAT3 controls ES cell self-renewal and pluripotency by a Myc-dependent mechanism. Development 132, 885–896 (2005).

    CAS  Article  PubMed  Google Scholar 

  13. Tomioka, M. et al. Identification of Sox-2 regulatory region which is under the control of Oct-3/4-Sox-2 complex. Nucleic Acids Res. 30, 3202–3213 (2002).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  14. Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006).

    CAS  PubMed  Article  Google Scholar 

  15. Takahashi, K., Okita, K., Nakagawa, M. & Yamanaka, S. Induction of pluripotent stem cells from fibroblast cultures. Nat. Protoc. 2, 3081–3089 (2007).

    CAS  PubMed  Article  Google Scholar 

  16. Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007).

    CAS  Article  PubMed  Google Scholar 

  17. Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917–1920 (2007).

    CAS  PubMed  Article  Google Scholar 

  18. Bilic, J. & Izpisua Belmonte, J.C. Concise review: induced pluripotent stem cells versus embryonic stem cells: close enough or yet too far apart? Stem Cells 30, 33–41 (2012).

    CAS  PubMed  Article  Google Scholar 

  19. Maherali, N. & Hochedlinger, K. Guidelines and techniques for the generation of induced pluripotent stem cells. Cell Stem Cell 3, 595–605 (2008).

    CAS  PubMed  Article  Google Scholar 

  20. Stadtfeld, M. et al. Aberrant silencing of imprinted genes on chromosome 12qF1 in mouse induced pluripotent stem cells. Nature 465, 175–181 (2010).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  21. Stadtfeld, M. et al. Ascorbic acid prevents loss of Dlk1-Dio3 imprinting and facilitates generation of all–iPS cell mice from terminally differentiated B cells. Nat. Genet. 44, 398–405 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  22. Okita, K., Ichisaka, T. & Yamanaka, S. Generation of germline-competent induced pluripotent stem cells. Nature 448, 313–317 (2007).

    CAS  Article  PubMed  Google Scholar 

  23. Wernig, M. et al. In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 448, 318–324 (2007).

    CAS  Article  PubMed  Google Scholar 

  24. Maherali, N. et al. Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution. Cell Stem Cell 1, 55–70 (2007).

    CAS  Article  PubMed  Google Scholar 

  25. Boland, M.J. et al. Adult mice generated from induced pluripotent stem cells. Nature 461, 91–94 (2009).

    CAS  Article  PubMed  Google Scholar 

  26. Kang, L., Wang, J., Zhang, Y., Kou, Z. & Gao, S. iPS cells can support full-term development of tetraploid blastocyst-complemented embryos. Cell Stem Cell 5, 135–138 (2009).

    CAS  Article  PubMed  Google Scholar 

  27. Zhao, X.Y. et al. iPS cells produce viable mice through tetraploid complementation. Nature 461, 86–90 (2009).

    CAS  Article  PubMed  Google Scholar 

  28. Miura, K. et al. Variation in the safety of induced pluripotent stem cell lines. Nat. Biotechnol. 27, 743–745 (2009).

    CAS  Article  PubMed  Google Scholar 

  29. Giorgetti, A. et al. Generation of induced pluripotent stem cells from human cord blood cells with only two factors: Oct4 and Sox2. Nat. Protoc. 5, 811–820 (2010).

    CAS  Article  PubMed  Google Scholar 

  30. Giorgetti, A. et al. Generation of induced pluripotent stem cells from human cord blood using OCT4 and SOX2. Cell Stem Cell 5, 353–357 (2009).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  31. Kim, J.B. et al. Oct4-induced pluripotency in adult neural stem cells. Cell 136, 411–419 (2009).

    CAS  PubMed  Article  Google Scholar 

  32. Kim, J.B. et al. Pluripotent stem cells induced from adult neural stem cells by reprogramming with two factors. Nature 454, 646–650 (2008).

    CAS  PubMed  Article  Google Scholar 

  33. Seki, T. et al. Generation of induced pluripotent stem cells from human terminally differentiated circulating T cells. Cell Stem Cell 7, 11–14 (2010).

    CAS  PubMed  Article  Google Scholar 

  34. Kim, J. et al. Reprogramming of postnatal neurons into induced pluripotent stem cells by defined factors. Stem Cells 29, 992–1000 (2011).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  35. Ruiz, S. et al. High-efficient generation of induced pluripotent stem cells from human astrocytes. PLoS One 5, e15526 (2010).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  36. Aasen, T. et al. Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes. Nat. Biotechnol. 26, 1276–1284 (2008).

    CAS  Article  PubMed  Google Scholar 

  37. Sommer, C.A. et al. Excision of reprogramming transgenes improves the differentiation potential of iPS cells generated with a single excisable vector. Stem Cells 28, 64–74 (2010).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  38. Soldner, F. et al. Parkinson's disease patient-derived induced pluripotent stem cells free of viral reprogramming factors. Cell 136, 964–977 (2009).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  39. Stadtfeld, M., Nagaya, M., Utikal, J., Weir, G. & Hochedlinger, K. Induced pluripotent stem cells generated without viral integration. Science 322, 945–949 (2008).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  40. Fusaki, N., Ban, H., Nishiyama, A., Saeki, K. & Hasegawa, M. Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome. Proc. Jpn. Acad. Ser. B Phys. Biol. Sci. 85, 348–362 (2009).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  41. Li, M. & Izpisua Belmonte, J.C. No factor left behind: generation of transgene-free induced pluripotent stem cells. Am. J. Stem Cells 1, 75–80 (2012).

    CAS  PubMed  Google Scholar 

  42. Jiang, J. et al. A core Klf circuitry regulates self-renewal of embryonic stem cells. Nat. Cell Biol. 10, 353–360 (2008).

    Article  CAS  PubMed  Google Scholar 

  43. Nakagawa, M., Takizawa, N., Narita, M., Ichisaka, T. & Yamanaka, S. Promotion of direct reprogramming by transformation-deficient Myc. Proc. Natl. Acad. Sci. USA 107, 14152–14157 (2010).

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  44. Nakagawa, M. et al. Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat. Biotechnol. 26, 101–106 (2008).

    CAS  PubMed  Article  Google Scholar 

  45. Dunn, S.J., Martello, G., Yordanov, B., Emmott, S. & Smith, A.G. Defining an essential transcription factor program for naive pluripotency. Science 344, 1156–1160 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  46. Feng, B. et al. Reprogramming of fibroblasts into induced pluripotent stem cells with orphan nuclear receptor Esrrb. Nat. Cell Biol. 11, 197–203 (2009).

    CAS  Article  PubMed  Google Scholar 

  47. Hou, P. et al. Pluripotent stem cells induced from mouse somatic cells by small-molecule compounds. Science 341, 651–654 (2013).

    CAS  Article  PubMed  Google Scholar 

  48. Ye, J. et al. Pluripotent stem cells induced from mouse neural stem cells and small intestinal epithelial cells by small molecule compounds. Cell Res. 26, 34–45 (2016).

    CAS  PubMed  Article  Google Scholar 

  49. Zhao, Y. et al. A XEN-like state bridges somatic cells to pluripotency during chemical reprogramming. Cell 163, 1678–1691 (2015).

    CAS  PubMed  Article  Google Scholar 

  50. Buganim, Y. et al. Single-cell expression analyses during cellular reprogramming reveal an early stochastic and a late hierarchic phase. Cell 150, 1209–1222 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  51. Polo, J.M. et al. A molecular roadmap of reprogramming somatic cells into iPS cells. Cell 151, 1617–1632 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  52. Ang, Y.S. et al. Wdr5 mediates self-renewal and reprogramming via the embryonic stem cell core transcriptional network. Cell 145, 183–197 (2011).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  53. Onder, T.T. et al. Chromatin-modifying enzymes as modulators of reprogramming. Nature 483, 598–602 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  54. Yu, C., Liu, K., Tang, S. & Ding, S. Chemical approaches to cell reprogramming. Curr. Opin. Genet. Dev. 28, 50–56 (2014).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  55. Ruiz, S. et al. A high proliferation rate is required for cell reprogramming and maintenance of human embryonic stem cell identity. Curr. Biol. 21, 45–52 (2011).

    CAS  PubMed  Article  Google Scholar 

  56. Li, H. et al. The Ink4/Arf locus is a barrier for iPS cell reprogramming. Nature 460, 1136–1139 (2009).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  57. Marión, R.M. et al. A p53-mediated DNA damage response limits reprogramming to ensure iPS cell genomic integrity. Nature 460, 1149–1153 (2009).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  58. Kawamura, T. et al. Linking the p53 tumour suppressor pathway to somatic cell reprogramming. Nature 460, 1140–1144 (2009).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  59. Müller, L.U. et al. Overcoming reprogramming resistance of Fanconi anemia cells. Blood 119, 5449–5457 (2012).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  60. Liu, G.H. et al. Modelling Fanconi anemia pathogenesis and therapeutics using integration-free patient-derived iPSCs. Nat. Commun. 5, 4330 (2014).

    CAS  PubMed  Article  Google Scholar 

  61. Dowey, S.N., Huang, X., Chou, B.K., Ye, Z. & Cheng, L. Generation of integration-free human induced pluripotent stem cells from postnatal blood mononuclear cells by plasmid vector expression. Nat. Protoc. 7, 2013–2021 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  62. Aasen, T. & Izpisua Belmonte, J.C. Isolation and cultivation of human keratinocytes from skin or plucked hair for the generation of induced pluripotent stem cells. Nat. Protoc. 5, 371–382 (2010).

    CAS  PubMed  Article  Google Scholar 

  63. Sancho-Martinez, I., Baek, S.H. & Izpisua Belmonte, J.C. Lineage conversion methodologies meet the reprogramming toolbox. Nat. Cell Biol. 14, 892–899 (2012).

    CAS  PubMed  Article  Google Scholar 

  64. Fatehullah, A., Tan, S.H. & Barker, N. Organoids as an in vitro model of human development and disease. Nat. Cell Biol. 18, 246–254 (2016).

    Article  CAS  PubMed  Google Scholar 

  65. Lian, X. et al. Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/b-catenin signaling under fully defined conditions. Nat. Protoc. 8, 162–175 (2013).

    CAS  PubMed  Article  Google Scholar 

  66. Shi, Y., Kirwan, P. & Livesey, F.J. Directed differentiation of human pluripotent stem cells to cerebral cortex neurons and neural networks. Nat. Protoc. 7, 1836–1846 (2012).

    CAS  Article  PubMed  Google Scholar 

  67. Ko, H.C. & Gelb, B.D. Concise review: drug discovery in the age of the induced pluripotent stem cell. Stem Cells Transl. Med. 3, 500–509 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  68. Gore, A. et al. Somatic coding mutations in human induced pluripotent stem cells. Nature 471, 63–67 (2011).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  69. Hussein, S.M. et al. Copy number variation and selection during reprogramming to pluripotency. Nature 471, 58–62 (2011).

    CAS  PubMed  Article  Google Scholar 

  70. Bhutani, K. et al. Whole-genome mutational burden analysis of three pluripotency induction methods. Nat. Commun. 7, 10536 (2016).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  71. Baira, E., Greshock, J., Coukos, G. & Zhang, L. Ultraconserved elements: genomics, function and disease. RNA Biol. 5, 132–134 (2008).

    CAS  PubMed  Article  Google Scholar 

  72. Li, M. & Izpisua Belmonte, J.C. Mending a faltering heart. Circ. Res. 118, 344–351 (2016).

    CAS  PubMed  Article  Google Scholar 

  73. Lancaster, M.A. et al. Cerebral organoids model human brain development and microcephaly. Nature 501, 373–379 (2013).

    CAS  Article  PubMed  Google Scholar 

  74. Zhong, X. et al. Generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs. Nat. Commun. 5, 4047 (2014).

    CAS  PubMed  Article  Google Scholar 

  75. McCracken, K.W. et al. Modelling human development and disease in pluripotent stem-cell-derived gastric organoids. Nature 516, 400–404 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  76. Sugii, S., Kida, Y., Berggren, W.T. & Evans, R.M. Feeder-dependent and feeder-independent iPS cell derivation from human and mouse adipose stem cells. Nat. Protoc. 6, 346–358 (2011).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  77. Panopoulos, A.D. et al. Rapid and highly efficient generation of induced pluripotent stem cells from human umbilical vein endothelial cells. PLoS One 6, e19743 (2011).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  78. Zhou, T. et al. Generation of human induced pluripotent stem cells from urine samples. Nat. Protoc. 7, 2080–2089 (2012).

    CAS  Article  PubMed  Google Scholar 

  79. Liu, H., Ye, Z., Kim, Y., Sharkis, S. & Jang, Y.Y. Generation of endoderm-derived human induced pluripotent stem cells from primary hepatocytes. Hepatology 51, 1810–1819 (2010).

    CAS  PubMed  Article  Google Scholar 

Download references

Acknowledgements

Work in the laboratory of J.C.I.B. was supported by the G. Harold and Leila Y. Mathers Charitable Foundation, the Leona M. and Harry B. Helmsley Charitable Trust (2012-PG-MED002), the Moxie Foundation, the NIH, Fundacion Dr. Pedro Guillen and UCAM.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Juan Carlos Izpisua Belmonte.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Li, M., Belmonte, J. Looking to the future following 10 years of induced pluripotent stem cell technologies. Nat Protoc 11, 1579–1585 (2016). https://doi.org/10.1038/nprot.2016.108

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2016.108

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

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