Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5⁺ stem cell

Nat Med. 2012 Mar 11;18(4):618-23. doi: 10.1038/nm.2695.


Adult stem-cell therapy holds promise for the treatment of gastrointestinal diseases. Here we describe methods for long-term expansion of colonic stem cells positive for leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5(+) cells) in culture. To test the transplantability of these cells, we reintroduced cultured GFP(+) colon organoids into superficially damaged mouse colon. The transplanted donor cells readily integrated into the mouse colon, covering the area that lacked epithelium as a result of the introduced damage in recipient mice. At 4 weeks after transplantation, the donor-derived cells constituted a single-layered epithelium, which formed self-renewing crypts that were functionally and histologically normal. Moreover, we observed long-term (>6 months) engraftment with transplantation of organoids derived from a single Lgr5(+) colon stem cell after extensive in vitro expansion. These data show the feasibility of colon stem-cell therapy based on the in vitro expansion of a single adult colonic stem cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells / metabolism*
  • Animals
  • Body Weight / physiology
  • Cells, Cultured
  • Chromogranin A / metabolism
  • Colon / cytology*
  • Colon / transplantation*
  • Cyclooxygenase 1 / metabolism
  • DNA-Binding Proteins / deficiency
  • Epithelium / physiology*
  • Green Fluorescent Proteins / genetics
  • In Vitro Techniques
  • Ki-67 Antigen / metabolism
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Transgenic
  • Organoids / cytology
  • Organoids / physiology
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism*
  • Rhodamines / metabolism
  • Time Factors
  • Transplants


  • Chromogranin A
  • DNA-Binding Proteins
  • Ki-67 Antigen
  • Lgr5 protein, mouse
  • Membrane Proteins
  • Rag2 protein, mouse
  • Receptors, G-Protein-Coupled
  • Rhodamines
  • chromogranin A, mouse
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • tetramethylrhodamine isothiocyanate
  • Cyclooxygenase 1
  • Ptgs1 protein, mouse