Human Intestinal Tissue with Adult Stem Cell Properties Derived from Pluripotent Stem Cells

Creators: Forster, Ryan; Chiba, Kunitoshi; Schaeffer, Lorian; Regalado, Samuel G.; Lai, Christine S.; Gao, Qing; Kiani, Samira; Farin, Henner F.; Clevers, Hans; Cost, Gregory J.; Chan, Andy; Rebar, Edward J.; Urnov, Fyodor D.; Gregory, Philip D.; Pachter, Lior; Jaenisch, Rudolf; Hockemeyer, Dirk

Abstract

Genetically engineered human pluripotent stem cells (hPSCs) have been proposed as a source for transplantation therapies and are rapidly becoming valuable tools for human disease modeling. However, many applications are limited due to the lack of robust differentiation paradigms that allow for the isolation of defined functional tissues. Here, using an endogenous LGR5-GFP reporter, we derived adult stem cells from hPSCs that gave rise to functional human intestinal tissue comprising all major cell types of the intestine. Histological and functional analyses revealed that such human organoid cultures could be derived with high purity and with a composition and morphology similar to those of cultures obtained from human biopsies. Importantly, hPSC-derived organoids responded to the canonical signaling pathways that control self-renewal and differentiation in the adult human intestinal stem cell compartment. This adult stem cell system provides a platform for studying human intestinal disease in vitro using genetically engineered hPSCs.

Additional Information

© 2014 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). Received 24 November 2013, Revised 4 May 2014, Accepted 5 May 2014, Available online 3 June 2014. We thank R. Alagappan, P. Xu, Dong Dong Wu, and Lei Zhang and the Sangamo Production group for expert technical assistance. We thank Frank Soldner, Thomas Sandmann, and Helen Bateup for helpful comments during the design of the experiment. We thank Nicki Watson from the Keck Imaging Facility at the Whitehead Institute for performing the EM analysis. R.F. is supported by the National Science Foundation Graduate Research Fellowship Program (GRFP) under grant DGE 1106400 and NIH training grant 2T32GM007232-36. K.C. was supported by a fellowship from the Nakajima Foundation. R.J. was supported by NIH grants R37-CA084198, RO1-CA087869, and RO1-HD045022, and by a grant from the HHMI. R.J. is an adviser to Stemgent and a cofounder of Fate Therapeutics. D.H. is a New Scholar in Aging of the Ellison Medical Foundation and is supported by the Glenn Foundation and the Shurl and Kay Curci Foundation. G.J.C., A.C., E.J.R., P.D.G., and F.D.U. are full-time employees of Sangamo BioSciences. Accession Numbers: The GEO accession number for the RNA-seq data reported in this paper is GSE56930.

Errata

Ryan Forster, Kunitoshi Chiba, Lorian Schaeffer, Samuel G. Regalado, Christine S. Lai, Qing Gao, Samira Kiani, Henner F. Farin, Hans Clevers, Gregory J. Cost, Andy Chan, Edward J. Rebar, Fyodor D. Urnov, Philip D. Gregory, Lior Pachter, Rudolf Jaenisch, Dirk Hockemeyer, Human Intestinal Tissue with Adult Stem Cell Properties Derived from Pluripotent Stem Cells, Stem Cell Reports, Volume 3, Issue 1, 8 July 2014, Page 215, ISSN 2213-6711, http://dx.doi.org/10.1016/j.stemcr.2014.06.014. (http://www.sciencedirect.com/science/article/pii/S2213671114002008)

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