Urothelium Tissue Engineering Using Bladder Mucosa From Transurethral Resection of Prostate
试验速览
- 阶段
- 不适用
- 状态
- 尚未招募
- 发起方
- 入组人数
- 365
- 主要终点
- Histological analysis of biopsy.
研究概览
简要总结
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.
详细描述
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).
研究设计
- 研究类型
- Observational
- 观察模型
- Other
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- Male
- 接受健康志愿者
- 否
入选标准
- •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
排除标准
- •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
结局指标
主要结局
Histological analysis of biopsy.
时间窗: 6 month
Histological analysis with standard coloration will assess the viability of urothelium. Signs of necrosis (ulcerations, destructions of urothelium structure) will be noted.
次要结局
- Association of the cultured cells with different scaffolds. Culture of cellularized scaffolds.(36 months)
- Histological analysis of cellularized scaffolds(36 months)
- Immunohistological analysis of biopsy.(6 month)
- Immunohistological analysis of cellularized scaffolds(36 months)
- RT-qPCR analysis of biopsy.(6 month)
- Histological analysis of the cultured cells.(12 months)
- Digestion of the biopsy and culture with adapted medium. Optimization of culture conditions.(12 months)
- Immunocytological analysis of the cultured cells.(12 months)
- RT-qPCR analysis of the cultured cells.(12 months)
- Development of original alginate freezed-dried scaffold(12 months)
- RT-qPCR analysis of cellularized scaffolds(36 months)
- Implantation of the cellularized scaffold in Nude mice(12 months)
- Histological analysis of implanted scaffolds(12 months)
- Immunohistological analysis of implanted scaffolds(12 months)
- Biophysical analysis of cellularized scaffolds(36 months)
- RT-qPCR analysis of implanted scaffolds(12 months)