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）：当正常膀胱细胞在培养物中生长时，它们保持正常，而当患病或异常膀胱细胞在培养物中生长时，它们保持异常。需要膀胱增大或再生的患者可能没有用于再生的健康膀胱细胞，因此需要替代的细胞来源。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，因此它们可能可用作组织工程膀胱再生的替代细胞来源。 BMSC通过骨髓抽吸获得，并且可以在体外扩增到足够的数量。它们不是干细胞，但显示出干细胞群的多种特征，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。本申请重点研究 BMSC 在膀胱再生方面的前景。第一个具体目标：(1) 表征 BMSC 与膀胱尿路上皮细胞 (UC) 体外共培养时的特征。 (1.1)分离并培养BMSC、UC和SMC，(1.2)用增强型绿色荧光蛋白(GFP)标记BMSC以供以后鉴定，(1.3)在体外将BMSC与UC在SIS上共培养。 （1.3.1）评估UC对BMSC细胞生长和密度的影响，（1.3.2）通过表型外观、分子分析和免疫组织化学染色评估在SIS上UC共培养中诱导BMSC分化的最佳时机。第二个具体目标：（2）确定BMSC在体内的存活情况及其对膀胱功能和再生的影响。 (2.1) 用增强的 GFP 标记 BMSC 以便以后识别，(2.2) 在体外 SIS 上共培养 GFP 标记的 BMSC 和 UC，(2.3) 在半囊切除术后将细胞支架结构植入犬膀胱中，(2.3.1) 分析膀胱使用尿动力学研究评估功能和容量，(2.3.2) 评估不同时间点的体内 BMSC 存活率，(2.3.3) 评估不同时间点的膀胱再生程度通过组织学、收缩性测定和免疫组织化学染色的时间点。