A perfusion bioreactor for intestinal tissue engineering.
BACKGROUND
Short gut syndrome is a devastating clinical problem with limited long-term treatment options. A unique characteristic of the normal intestinal epithelium is its capacity for regeneration and adaptation. Despite this tremendous capacity in vivo, one of the major limitations in advancing the understanding of intestinal epithelial differentiation and proliferation has been the difficulty in maintaining primary cultures of normal gut epithelium in vitro. A perfusion bioreactor system has been shown to be beneficial in long-term culture and bioengineering of a variety of tissues. The purpose of this study is to design and fabricate a perfusion bioreactor for intestinal tissue engineering.


METHODS
A perfusion bioreactor is fabricated using specific parameters. Intestinal epithelial organoid units harvested from neonatal rats are seeded onto biodegradable polymer scaffolds and cultured for 2 d in the bioreactor. Cell attachment, viability, and survival are assessed using MTT assay, scanning electron micrograph, and histology.


RESULTS
A functional perfusion bioreactor was successfully designed and manufactured. MTT assay and scanning electron micrograph demonstrated successful attachment of viable cells onto the polymer scaffolds. Histology confirmed the survival of intestinal epithelial cells seeded on the scaffolds and cultured in the perfusion bioreactor for 2 days.


CONCLUSIONS
A functional perfusion bioreactor can be successfully fabricated for the in-vitro cultivation of intestinal epithelial cells. With further optimization, the perfusion bioreactor may be a useful in in-vitro system for engineering new intestinal tissue.
Kim
Stephen S
SS
Seattle Children's Hospital Research Institute, Children's Hospital and Regional Medical Center, Seattle, Washington 98105, USA. stephen.kim@seattlechildrens.org
Penkala
Rebecca
R
Abrahimi
Parwiz
P
D000818:Animals; D000831:Animals, Newborn; D019149:Bioreactors; D002448:Cell Adhesion; D002470:Cell Survival; D004847:Epithelial Cells / Q000166:cytology; D004867:Equipment Design; D007422:Intestines / Q000166:cytology* / Q000502:physiology*; D010477:Perfusion; D011108:Polymers; D051381:Rats; D011917:Rats, Inbred Lew; D012038:Regeneration; D012778:Short Bowel Syndrome; D023822:Tissue Engineering / Q000295:instrumentation*
D011108:Polymers