Protocol

Organ Regeneration

Volume 1001 of the series Methods in Molecular Biology pp 299-309

Date:

A “Living Bioreactor” for the Production of Tissue-Engineered Small Intestine

  • Daniel E. LevinAffiliated withDivision of Pediatric Surgery, Saban Research Institute, Children’s Hospital Los Angeles, Keck School of Medicine of the University of Southern California
  • , Frederic G. SalaAffiliated withDivision of Pediatric Surgery, Saban Research Institute, Children’s Hospital Los Angeles, Keck School of Medicine of the University of Southern California
  • , Erik R. BarthelAffiliated withDivision of Pediatric Surgery, Saban Research Institute, Children’s Hospital Los Angeles, Keck School of Medicine of the University of Southern California
  • , Allison L. SpeerAffiliated withDivision of Pediatric Surgery, Saban Research Institute, Children’s Hospital Los Angeles, Keck School of Medicine of the University of Southern California
  • , Xiaogang HouAffiliated withDivision of Pediatric Surgery, Saban Research Institute, Children’s Hospital Los Angeles, Keck School of Medicine of the University of Southern California
  • , Yasuhiro TorashimaAffiliated withDivision of Pediatric Surgery, Saban Research Institute, Children’s Hospital Los Angeles, Keck School of Medicine of the University of Southern California
  • , Tracy C. GrikscheitAffiliated withDivision of Pediatric Surgery, Saban Research Institute, Children’s Hospital Los Angeles, Keck School of Medicine of the University of Southern California

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Here, we describe the use of a mouse model as a living bioreactor for the generation of tissue-engineered small intestine. Small intestine is harvested from donor mice with subsequent isolation of organoid units (a cluster of mesenchymal and epithelial cells). Some of these organoid units contain pluripotent stem cells with a preserved relationship with the mesenchymal stem cell niche. A preparation of organoid units is seeded onto a biodegradable scaffold and implanted intraperitoneally within the omentum of the host animal. The cells are nourished initially via imbibition until neovascularization occurs. This technique allows the growth of fully differentiated epithelium (composed of Paneth cells, goblet cells, enterocytes and enteroendocrine cells), muscle, nerve, and blood vessels of donor origin. Variations of this technique have been used to generate tissue-engineered stomach, large intestine, and esophagus. The variations include harvest technique, length of digestion, and harvest times.

Key words

Tissue engineering Small intestine Organoid unit Short bowel syndrome Living bioreactor