In vitro tissue engineering of smooth muscle sheets with peristalsis using a murine induced pluripotent stem cell line Atsushi Yoshida a , Kenny Chitcholtan b , John J. Evans b,c , Volker Nock c , Spencer W. Beasley a, ⁎ a Department of Paediatric Surgery, Christchurch School of Medicine and Health Sciences, Christchurch Hospital, Private Bag 4710, Christchurch, New Zealand b Laboratory for Cell and Protein Regulation, Department of Obstetrics and Gynaecology, University of Otago, Christchurch Hospital, Christchurch, New Zealand c MacDiarmid Institute for Advanced Materials and Nanotechnology, Department of Electrical and Computer Engineering, University of Canterbury, Private Bag 4800, Christchurch, New Zealand Received 2 November 2011; accepted 10 November 2011 Key words: Short gut; Smooth muscle; Induced pleuripotent stem cells (iPS); Tissue engineering Abstract Purpose: Currently, 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. Method: Induced 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. Results: Outgrowth 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. Conclusion: Tissue 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. © 2012 Elsevier Inc. All rights reserved. Short bowel syndrome, a state of malabsorption that may necessitate prolonged parenteral nutrition [1], can occur after extensive small bowel resection for a variety of conditions including midgut volvulus, gastroschisis, necrotizing enter- colitis, mesenteric ischemia, and inflammatory bowel disease. Therapeutic options are limited, and none is entirely Presented at the 58th Annual Meeting of the British Association of Paediatric Surgeons, Belfast, Northern Ireland, July 20-22, 2011. ⁎ Corresponding author. Tel.: +64 33 640432; fax: +64 33 641584. E-mail address: spencer.beasley@cdhb.govt.nz (S.W. Beasley). www.elsevier.com/locate/jpedsurg 0022-3468/$ – see front matter © 2012 Elsevier Inc. All rights reserved. doi:10.1016/j.jpedsurg.2011.11.027 Journal of Pediatric Surgery (2012) 47, 329–335