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1 Division of Gastroenterology, University Hospital, Nottingham NG7 2UH, United Kingdom; and 2 Gastrointestinal Unit, Massachusetts General Hospital, Boston, Massachusetts 02114-2696
-After injury
and loss of epithelial cells, intestinal barrier function is
reestablished by migration of viable epithelial cells from the wound
edge (restitution). Myofibroblasts are located close to the basal
surface of epithelial cells. This study aimed to investigate the role
of human colonic subepithelial myofibroblasts in epithelial
restitution. Primary cultures of subepithelial myofibroblasts were
established. Monolayers of the epithelial cell lines IEC-6 and T84 were
"wounded" in a standard manner to create an in vitro model of
restitution. Migration of epithelial cells across the wound edge was
assessed following culture in myofibroblast-conditioned medium.
Myofibroblast expression of transforming growth factor (TGF)-
isoforms was examined using RT-PCR, and TGF-
isoform bioactivity was assessed using Mv 1 Lu bioassay.
Myofibroblast-conditioned medium, via a TGF-
-dependent pathway,
significantly enhanced migration of epithelial cells across the wound
edge and significantly inhibited cell proliferation in wounded
monolayers. Messenger RNA for TGF-
1, -
2, and -
3 was detected
in the myofibroblasts, and Mv 1 Lu bioassay showed the presence of
predominantly bioactive TGF-
3. This study shows that human colonic
subepithelial myofibroblasts secrete predominantly bioactive TGF-
3
and enhance restitution in wounded epithelial monolayers
via a TGF-
-dependent pathway.
transforming growth factor-
; wound repair
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