Combined injection of three different lineages of early-differentiating human amniotic fluid-derived cells restores urethral sphincter function in urinary incontinence.
OBJECTIVE
To investigate whether a triple combination of early-differentiated cells derived from human amniotic fluid stem cells (hAFSCs) would show synergistic effects in urethral sphincter regeneration.


METHODS
We early-differentiated hAFSCs into muscle, neuron and endothelial progenitor cells and then injected them into the urethral sphincter region of pudendal neurectomized ICR mice, as single-cell, double-cell or triple-cell combinations. Urodynamic studies and histological, immunohistochemical and molecular analyses were performed.


RESULTS
Urodynamic study showed significantly improved leak point pressure in the triple-cell-combination group compared with the single-cell- or double-cell-combination groups. These functional results were confirmed by histological and immunohistochemical analyses, as evidenced by the formation of new striated muscle fibres and neuromuscular junctions at the cell injection site. Molecular analysis showed higher target marker expression in the retrieved urethral tissue of the triple-cell-combination group. The injection of early-differentiated hAFSCs suppressed in vivo host CD8 lymphocyte aggregations and did not form teratoma. The nanoparticle-labelled early-differentiated hAFSCs could be tracked in vivo with optical imaging for up to 14 days after injection.


CONCLUSIONS
Our novel concept of triple-combined early-differentiated cell therapy for the damaged sphincter may provide a viable option for incontinence treatment.
Joint Institute for Regenerative Medicine, Kyungpook National University Hospital, Daegu, Korea.
So Young
SY
Chun
Joon Beom
JB
Kwon
Seon Yeong
SY
Chae
Jong Kil
JK
Lee
Jae-Sung
JS
Bae
Bum Soo
BS
Kim
Hyun Tae
HT
Kim
Eun Sang
ES
Yoo
Jeong Ok
JO
Lim
James J
JJ
Yoo
Wun-Jae
WJ
Kim
Bup Wan
BW
Kim
Tae Gyun
TG
Kwon
D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D058948:Cell Tracking; D005260:Female; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000166:cytology*; D001743:Urinary Bladder / Q000166:cytology*; D014549:Urinary Incontinence / Q000628:therapy*
amniotic fluid; regeneration; stem cells; urethral sphincter; urinary incontinence