In vitro tissue engineering of smooth muscle sheets with peristalsis using a murine induced pluripotent stem cell line

PurposeCurrently, therapeutic options for short gut syndrome are limited, ranging from total parenteral nutrition pending intestinal adaption to small bowel transplantation with its associated problems of rejection. The aim of this study was to differentiate murine induced pluripotent stem cells (iPSCs) into gut tissues, including contracting smooth muscle, as a precursor to using tissue engineering techniques to generate functioning intestinal tissue from the patient's own cells.MethodInduced pluripotent stem cells were cultured for 6 days in low adherent 96-well plates (500-5000 cells per well) in a differentiation medium comprising Knockout Dulbecco's modified Eagle medium (Invitrogen, Carlsbad, Calif) supplemented with 10% fetal bovine serum, l-glutamine, nonessential amino acids, 2-mercaptoethanol, penicillin, and streptomycin. After 6 days, the embryoid body that had formed in each well was transferred into a 12-well plate, each well containing 8 embryoid bodies.ResultsOutgrowth culture produced differentiated cell clusters, including cardiaclike cells, sheets of smooth muscle with peristalticlike activity, and mucosal cells. Cardiaclike cells exhibited spontaneous rhythmic regular contraction, but the sheets of smooth muscle contracted in a co-ordinated way, just as is seen in peristaltic bowel, as shown using video imaging.ConclusionTissue engineering techniques using iPSCs have the potential to ultimately replace the need for small bowel transplantation and allow patients to “grow” their own small bowel.