Brief Communication | Published:

Isolation and in vitro expansion of human colonic stem cells

Nature Medicine volume 17, pages 12251227 (2011) | Download Citation

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Abstract

Here we describe the isolation of stem cells of the human colonic epithelium. Differential cell surface abundance of ephrin type-B receptor 2 (EPHB2) allows the purification of different cell types from human colon mucosa biopsies. The highest EPHB2 surface levels correspond to epithelial colonic cells with the longest telomeres and elevated expression of intestinal stem cell (ISC) marker genes. Moreover, using culturing conditions that recreate the ISC niche, a substantial proportion of EPHB2-high cells can be expanded in vitro as an undifferentiated and multipotent population.

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Change history

  • 20 September 2011

     In the version of this article initially published online, the author list was missing the name Herbert Auer, and the name of Mercedes Gallardo was incorrectly given as María M. Gallardo. The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank Genentech for the EphB2-specific antibody Mab 2H9 and the Institute for Research in Biomedicine transcriptomic facility, electronic microscopy service (Universitat de Barcelona) and Cell separation unit (UB) for expert assistance. We thank L. Lopez Vilaro and E. Calderón Gómez for help with mucosas. This work is supported by grants to E.B. from the CONSOLIDER and Plan Nacional I+D+i Programs from Spanish Ministry of Science and Innovation (MICINN) and the European Research Council (ERC-FP7). Work was furthermore supported by grants from Instituto de Salud Carlos III FEDER (RD09/0076/00036). The 'Xarxa de Bancs de tumors' is sponsored by Pla Director d'Oncologia de Catalunya (XBTC). The M.A.B. laboratory is founded by the MICINN, the EU, the Botin and Lilly Foundations, and the Korber European Science Award to M.A.B. P.J. holds a Marie Curie Fellowship (FP7).

Author information

Author notes

    • Peter Jung
    •  & Toshiro Sato

    These authors contributed equally to this work.

Affiliations

  1. Oncology program, Institute for Research in Biomedicine, Barcelona, Spain.

    • Peter Jung
    • , Anna Merlos-Suárez
    • , Francisco M Barriga
    • , Elena Sancho
    •  & Eduard Batlle
  2. Hubrecht Institute and University Medical Center Utrecht, Utrecht, The Netherlands.

    • Toshiro Sato
    •  & Hans Clevers
  3. Department of Gastroenterology, School of Medicine, Keio University, Shinjuku-ku, Tokyo, Japan.

    • Toshiro Sato
  4. Department of Pathology, Hospital Universitari del Mar, Universitat Autònoma de Barcelona, Barcelona, Spain.

    • Mar Iglesias
  5. Biostatistics and Bioinformatics Unit, Institute for Research in Biomedicine, Barcelona, Spain.

    • David Rossell
  6. Functional Genomics Core Facility, Institute for Research in Biomedicine, Barcelona, Spain.

    • Herbert Auer
  7. Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Research Center, Madrid, Spain.

    • Mercedes Gallardo
    •  & Maria A Blasco
  8. Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain.

    • Eduard Batlle

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Contributions

E.B. and P.J., study concept and design and writing of the manuscript. T.S. and H.C., development of original human colon culture method. P.J., acquisition, analysis and interpretation of data and modifications of the human CoSC culture protocol. A.M.-S. and F.M.B., profiling of human colon stem cells. D.R., bioinformatic analysis. E.S., conceptual and logistic support. M.M.G. and M.A.B., telomere length measurements. M.I., sample preparation and development of EphB2 sorting method.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Eduard Batlle.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–11, Supplementary Data, Supplementary Table 1 and Supplementary Methods

Excel files

  1. 1.

    Supplementary Table 2

    List of genes whose expression is enriched in EPHB2-high cells.

  2. 2.

    Supplementary Table 3

    Genome-wide expression analysis of EPHB2-high, EPHB2-medium, EPHB2-low and EPHB2-negative cells from 3 independent colon mucosa samples.

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DOI

https://doi.org/10.1038/nm.2470

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