Massachusetts Chapter of the American College of Surgeons

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Donor Mesenchymal Stem Cell Kinetics in Transamniotic Stem Cell Therapy for Experimental Gastroschisis
Alexander V Chalphin; Sarah A Tracy; Ina Kycia; Christopher Chan; Adam Finkelstein; David Zurakowski; Dario O Fauza
Boston Children's Hospital, Boston, MA, USA

Background: Transamniotic stem cell therapy (TRASCET) with mesenchymal stem cells has been shown to minimize damage to the exposed bowel in animal models of gastroschisis. Donor cell routing in this setting, however, remained unknown. We sought to comprehensively scrutinize donor cell kinetics after TRASCET in the rodent model of gastroschisis.

Methods: A gastroschisis was surgically created in 102 rat fetuses at gestation day 18 (term=22 days), immediately followed by volume-matched amniotic injections of either amniotic fluid mesenchymal stem cells (afMSCs) labeled with a luciferase reporter gene (n=58), or luciferase protein alone (n=44). Infused afMSCs consisted of syngeneic rat cells with mesenchymal progenitor identity confirmed by flow cytometry. Samples from twelve anatomical sites (placenta, umbilical cord, skin, brain, heart, lung, liver, spleen, kidney, bone marrow, and bowel) from survivors were screened for luciferase activity via microplate luminometry at term. Statistical analysis included Mann-Whitney U-test, Wald test, and kappa coefficient (p<0.05).

Results: Overall survival was 42% (43/102), with no significant difference between the two groups (p=0.82). When controlled by acellular luciferase, donor afMSCs were identified selectively in the placenta (p<0.001) and bowel (p=0.005), independently of the dams (respectively, p<0.001 and p=0.041). Bowel homing was documented exclusively in areas exposed to the amniotic cavity. There was no mutual correlation between placental and bowel homing (kappa=-0.02; p=0.91).

Conclusions: afMSCs home to specific sites after TRASCET in the setting of gastroschisis. Placental and intestinal homing are fundamental yet seemingly independent constituents of cell trafficking, suggesting that both direct amniotic seeding and hematogenous routing take place.

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