Spina Bifida Clinical Trial
Official title:
Urothelium Tissue Engineering Using Bladder Mucosa From Transurethral Resection of Prostate
Different clinical conditions can require urinary bladder augmentation or replacement. Tissue
engineered bladder has been clinically evaluated but is not recommended due to diverse side
effects. Thus, there is a real interest for the development of regenerative approach with
innovative scaffolds and cell transplantation.
The investigators propose the use of urothelial cells obtained by Trans-Urethral Resection of
Prostate or bladder (TURP) to obtain a tissue engineered urothelium in association with
different scaffolds.
Bladder biopsies will be obtained during cystoscopy, conserved in culture medium (DMEM®),
digested by dispase and sowed on collagen-coated culture support. Keratinocyte Serum Free
Medium (KSFM) will be used for proliferation. Microscopy, immunohistochemistry, RNA
extraction, Reverse Transcription and quantitative Polymerase Chain Reaction (RT-qPCR) will
be performed during passages. Cell culture conditions will be optimized to improve
proliferation and avoid loss of differentiation. The investigators will develop scaffolds
based on sodium alginate hydrogels, followed by freeze-drying to generate porous sponges (at
-20°C and -80°C). Cultured cells will be associated to these original scaffolds and to other
scaffolds, for example alginate hydrogels or Collagen Cell Carrier (CCC), cultivated for 28
days and analyzed. Histological and immunohistological appearance of cellularized scaffolds
will be compared to assess the effectiveness of each scaffold for tissue engineering in
urothelium.
Cellularized scaffolds will be studied in vitro (Transepithelial Electrical Resistance,
impermeability, ability to be stitched, resistance to urine) and in vivo in ectopic location
(subcutaneous location in Nude mice) or in orthotopic location (bladder augmentation in small
animal).
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