Bone Marrow Stromal Cells for Bladder Tissue Engineering

用于膀胱组织工程的骨髓基质细胞

• 批准号: 7174575
• 负责人: YUANYUAN no ZHANG
• 金额: $ 17.94万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7174575
• 关键词: biotechnology; cell differentiation; dogs; hematopoietic stem cells; immunocytochemistry; regeneration; smooth muscle; tissue /cell culture; tissue engineering; urinary bladder; vascular endothelium

DESCRIPTION (provided by applicant): When normal bladder cells are grown in culture they remain normal, and when diseased or abnormal bladder cells are grown in culture they remain abnormal. A patient needing bladder augmentation or regeneration likely does not have healthy bladder cells for regeneration, and therefore needs an alternative cell source. Because Bone Marrow Stromal Cells (BMSC) have the capacity to be cultured and differentiated into smooth muscle-like cells, they may be useful as an alternative cell source for tissue engineered bladder regeneration. BMSC are obtained by bone marrow aspiration and can be expanded to sufficient numbers in vitro. They are not stem cells, but display multiple features of a stem cell population, including similar cell proliferation, histological appearance, and contractile phenotype compared to primary smooth muscle cell (SMC) cultures. This application focuses on researching the promising potential of BMSC for bladder regeneration. First Specific Aim: (1) Characterize BMSC when co-cultured with bladder urothelial cells (UC) in vitro. (1.1) Isolate and culture BMSC, UC, and SMC, (1.2) Label BMSC with enhanced Green Fluorescent Protein (GFP) for later identification, (1.3) Co-culture BMSC with UC on SIS in vitro. (1.3.1) Assess effect of UC on BMSC cell growth and density, (1.3.2) Evaluate optimal timing to induce BMSC differentiation in UC co-culture on SIS through phenotypic appearance, molecular analysis, and immunohistochemical staining. Second Specific Aim: (2) Determine the in vivo survival of BMSC and their impact on bladder function and regeneration. (2.1) Label BMSC with enhanced GFP for later identification, (2.2) Co-culture GFP labeled BMSC and UC on SIS in vitro, (2.3) Implant cell-scaffold construct into a canine bladder after hemicystectomy, (2.3.1) Analyze bladder function and capacity using urodynamic studies, (2.3.2) Evaluate in vivo BMSC survival at different time points, (2.3.3) Evaluate extent of bladder regeneration at different time points through histology, contractility assays, and immunohistochemical staining.

描述（由申请人提供）：当正常的膀胱细胞在培养物中生长时，它们保持正常，并且在培养物中生长或异常的膀胱细胞时，它们仍然异常。需要增强膀胱或再生的患者可能没有健康的膀胱细胞进行再生，因此需要替代细胞来源。由于骨髓基质细胞（BMSC）具有培养和分化为平滑肌样细胞的能力，因此它们可能是组织工程膀胱再生的替代细胞来源。 BMSC是通过骨髓抽吸获得的，可以在体外扩展到足够的数量。它们不是干细胞，而是显示干细胞群体的多种特征，包括与原代平滑肌细胞（SMC）培养物相比，相似的细胞增殖，组织学外观和收缩表型。该应用的重点是研究BMSC在膀胱再生中的有希望的潜力。第一个特定目的：（1）在体外与膀胱尿皮细胞（UC）共培养时，表征BMSC。 （1.1）分离株和培养的BMSC，UC和SMC，（1.2）标记BMSC，具有增强的绿色荧光蛋白（GFP），可用于以后鉴定，（1.3）共培养BMSC与SIS在体外的UC。 （1.3.1）评估UC对BMSC细胞生长和密度的影响，（1.3.2）评估通过表型外观，分子分析和免疫组织化学染色来诱导UC共培养中BMSC分化的最佳时机。第二个特定目的：（2）确定BMSC的体内存活及其对膀胱功能和再生的影响。 （2.1）具有增强GFP的标签BMSC，以供以后鉴定，（2.2）共培养GFP在体外SIS上标记为BMSC和UC，（2.3）植入细胞连接构建体后，将血清结肠切除术后的犬膀胱构造成犬类膀胱，（2.3.1）（2.3.1）分析膀胱功能和使用URDORDYNAMIC研究（2.2.2.2.2），VIV（2.3.2） （2.3.3）通过组织学，收缩力分析和免疫组织化学染色评估不同时间点膀胱再生的程度。