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Tissue Engineering Approaches For Treating Long Gap Esophageal Atresia
*Ishna Sharma1, *Todd Jensen1, *Adam Mitchell1, *Wael Sayej2, Christine Finck2
1Uconn Health, Farmington, CT; 2Connecticut Children's Medical Center, Hartford, CT

Objective:
Esophageal atresia occurs in 1 in 3000 births. Typically, surgical repair includes reconnection of the esophagus or in cases were the esophagus cannot be reconnected, interposition of a piece of stomach or intestine. These surgical options cause significant morbidity, therefore, a new therapeutic option is needed. The ability to obtain cells prior to birth and utilize those cells to augment a developmental defect would be novel.
Design:
Porcine amniotic fluid stem cells were obtained via amniocentesis approximately
70-90 days prior to term. Cells were isolated, expanded and characterized via flow cytometry and qRT-PCR. Cells were over 90% positive for mesenchymal stem cell markers CD73, CD90 and CD105. Approximately 10 million cells were seeded onto the extra luminal surface of a polyurethane scaffold and allowed to incubate for 7 days in a rotating hollow organ bioreactor. Approximately 4cm of intra-thoracic esophagus was removed in 2 piglets from the same litter as the isolated amniotic fluid stem cells and replaced with the seeded scaffolding in accordance with IACUC approval (HH#2014-0132). The lumen was stented open with a 10 mm biliary stent.
Results:
The stent was removed and changed at 3 weeks, which revealed that the scaffold had been extruded and the lumen was regenerated. Piglets were fed via feeding tube for 5-6 weeks, and then successfully transitioned to oral feeding. These animals were allowed to grow for 6 months prior to harvest and analysis.
Conclusions:
These piglets demonstrate the translational application of maternal amniotic fluid stem cells and a bioengineered synthetic scaffold in replacing missing gaps of esophageal tissue in offspring. Further studies are ongoing to delineate the mechanism of esophageal regeneration.


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