Korean J Urol.  2009 Mar;50(3):282-287.

Preliminary Study of Tissue-Engineered Ileal Conduit Using Poly (epsilon-Caprolactone) (PCL) Nano-Sheet Seeded with Muscle-Derived Stem Cells

• Lee YS
• Cho SY
• Kim HW
• Kang SH
• Kim HY
• Lee JY
• Kwon EB
• Hong SK
• Lee JY

Abstract

PURPOSE: We studied the feasibility and results of a tissue-engineered ileal conduit using a poly (epsilon-caprolactone) (PCL) nano-sheet seeded with muscle-derived stem cells to replace a conventional ileal conduit in rats.
MATERIALS AND METHODS
Muscle-derived stem cells were isolated from the gastrocnemius muscle of female Sprague-Dawley rats (200-250 g, n=6) by use of a preplate technique and were cultured on a PCL nano-sheet. The PCL nano-sheet was implanted into the omentum of rats and was then made into a conical shaped conduit. Rats were sacrificed 4 and 8 weeks after implantation, and morphologic changes were assessed by H&E and immunofluorescence staining, including DAPI staining and staining for myogenin and myosin heavy chain (MyHC).
RESULTS
All rats survived until the end of the experiment. A minimal inflammatory reaction was observed around the PCL nano-sheet in the 4 week specimens but was found to be reduced in the 8 week specimen. Muscle bundles were identified at week 4 as well as week 8 after implantation on H&E staining. Around the PCL sheet, immunostaining for both myogenin and MyHC were positive, indicating skeletal muscle differentiation and ingrowth into the PCL sheet.
CONCLUSIONS
A PCL nano-sheet seeded with muscle-derived stem cells showed successful skeletal muscle differentiation at 4 and 8 weeks after implantation. This preliminary result supports the feasibility of a tissue-engineered ileal conduit using a PCL nano-sheet (seeded with muscle-derived stem cells) in place of conventional ileal conduits.

Keyword

Urinary diversion; Tissue engineering; Stem cells

MeSH Terms

Animals
Female
Fluorescent Antibody Technique
Humans
Hypogonadism
Indoles
Mitochondrial Diseases
Muscle, Skeletal
Muscles
Myogenin
Myosin Heavy Chains
Omentum
Ophthalmoplegia
Rats
Rats, Sprague-Dawley
Seeds
Stem Cells
Tissue Engineering
Urinary Diversion
Hypogonadism
Indoles
Mitochondrial Diseases
Myogenin
Myosin Heavy Chains
Ophthalmoplegia

Figure

• 

  Fig. 1. Diagram of the poly (ε-caprolac-tone) (PCL) nano-sheet conduit surgical procedure. The PCL nano-sheet was attached to the omentum and made into a conical shape. The conical base of the conduit was opened for outflow of urine. Both ureters were dissected and mobilized level of renal hilum and ureteroneocystostomy were done. The conical base of the PCL nano-sheet conduit was secured to the rat’s abdominal skin.

• 

  Fig. 2. Histologic aspects of the muscle fibers on the poly (ε-caprolactone) (PCL) nano-sheet in the mesentery at 4 weeks after implantation. (A) Muscular bundles on the PCL nano-sheet can be seen (H&E, x100). (B) Neutrophils and lymphocytes can be seen around the PCL nano-sheet (H&E, x400). (C) PKH26 immunostaining shows differentiated skeletal muscle cells (red color) on the PCL nano-sheet and omental tissue after cryosection (x100). (D) DAPI immunostaining shows positive cells surrounding the sheet after cryosection (x100). (E) Immunostaining of myogenin and myosin heavy chain shows skeletal muscle bundle fibers (green color, x100).

• 

  Fig. 3. Histologic aspects of the muscle fibers on the poly (ε -caprolactone) (PCL) nano-sheet in the mesentery at 8 weeks after implantation. (A) Increased muscular bundles on the PCL nano-sheet can be seen (H&E, x100). (B) Decreased inflammatory cells around the PCL nano-sheet (H&E, x400). (C) PKH26 immunostaining shows the spread of differentiated skeletal muscle cells (red color) on the PCL nano-sheet and omental tissue after cryosection (x100). (D) DAPI immunostaining shows increased positive cells surrounding the sheet after cryosection (x100). (E) Immunostaining of myogenin and myosin heavy chain shows increased various types of cells and thick skeletal muscle bundle fibers (green color, x100).

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