Urothelium Tissue Engineering Using Biopsies From Transurethral Resection of Prostate
Urothelium Tissue Engineering Using Biopsies 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).
Inclusion Criteria:

 -  Patient needing a Transurethral Resection of Prostate (TURP)

 -  Weight of prostate (evaluated by ultrasonography)greater than or equal to 30 grams

 -  Affiliation to a social security system

 -  Patient over the age of majority

 -  Patient receiving complete information on research organization without opposition to
 the use of biological specimen

 Exclusion Criteria:

 -  Patient for whom no TURP is realized during endoscopic procedure

 -  Patient for whom no resection is realized on the bladder neck

 -  Patient with prostate weight estimated under 30 grams

 -  Patient who opposed to the realization of the study