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Updated: Oct 28, 2026
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Published on: July 27, 2014
Human amniotic fluid stem cell-derived extracellular vesicles enhance enteric neural crest cell extension following
Nana Nakazawa-Tanaka1, Naho Fujiwara2, Katsumi Miyahara3
1Department of Pediatric Surgery, Juntendo University Urayasu Hospital, 2-1-1 Tomioka, Urayasu, Chiba, 279-0021, Japan. nana-n@juntendo.ac.jp.
Purpose:
Enteric neural crest-derived cell (ENCC) transplantation is a potential regenerative strategy for Hirschsprung disease (HSCR), but approaches to enhance transplanted ENCC extension remain limited. We investigated whether human amniotic fluid stem cell-derived extracellular vesicles (hAFSC-EVs) enhance ENCC extension in aganglionic gut.
Methods:
Neurospheres (NS) generated from embryonic day 18.5 (E18.5) Sox10-Venus+ ENCCs were transplanted into aganglionic gut from E12.5 Ednrb knockout mice and cultured with (EV+, n = 4) or without (EV-, n = 4) 1% hAFSC-EVs. ENCC extension was assessed over 72 h, and explants were examined by immunofluorescence staining. RNA sequencing was performed on Sox10-Venus+ ENCC-derived NS cultured with or without 1% hAFSC-EVs.
Results:
At 72 h, maximum ENCC extension was greater in EV+ than EV- explants (483.9 ± 55.7 vs. 303.2 ± 119.9 μm, P = 0.049), as was extension area (45,546 ± 18,623 vs. 15,049 ± 11,510 μm², P = 0.039). Ki67-positive nuclei in close association with Sox10-Venus- and TUJ1-positive structures were more readily observed in EV-treated explants. RNA sequencing demonstrated upregulation of oxidative phosphorylation- and cell-cycle-related genes.
Conclusion:
hAFSC-EVs enhanced transplanted ENCC extension in aganglionic gut, accompanied by transcriptional changes in bioenergetic and proliferative pathways. These findings suggest that hAFSC-EVs may provide a strategy to enhance ENCC-based regenerative approaches for HSCR.
