Urothelium Tissue Engineering Using Biopsies From Transurethral Resection of Prostate

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).

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