Enhancing Cell Survival on Aged Bruch's Membrane

增强老化布鲁赫膜上的细胞存活率

基本信息

批准号：
7248593
负责人：
MARCO A ZARBIN
金额：
$ 15.1万
依托单位：
UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-05-01 至 2008-04-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Resurfacing of diseased or iatrogenically damaged Bruch's membrane with healthy retinal pigment epithelium (RPE) has been proposed as adjunctive treatment for age-related macular degeneration (AMD). However, aged submacular Bruch's membrane does not support transplanted RPE survival and differentiation. With aging and AMD, Bruch's membrane undergoes numerous changes that could affect ligand availability. These observations lead to the following hypothesis: Aging changes mask extracellular matrix ligand availability to transplanted RPE cells, impairing post-attachment events and leading, in turn, to cell death or inability of the cells to differentiate. The following specific aims will determine whether cell health can be improved by resurfacing Bruch's membrane with bovine corneal endothelial cell extracellular matrix (BCE-ECM), a matrix that supports RPE growth and differentiation. Aim 1: Determine whether bovine corneal endothelial cells (BCE) can "resurface" Bruch's membrane. Aim 2: Determine whether cultured RPE cells can form focal adhesions on BCE-ECM-coated RPE basement membrane/Bruch's membrane and whether short-term cell survival is improved. Aim 3: Determine whether RPE cell longer-term survival is improved on BCE-ECM-coated Bruch's membrane. These studies will utilize an organ explant culture system of aged, human submacular Bruch's membrane. Resurfacing will be accomplished by culturing BCE on explants for a period of time to allow BCE-ECM deposition, followed by removal of cells and seeding with cultured RPE cells. Explants will be analyzed with scanning, light, and confocal microscopy. If RPE survival and differentiation are enhanced via this approach, the studies will lay the foundation for further studies using ligand components of BCE-ECM, which could form a basis for clinical treatment of Bruch's membrane to enhance RPE graft survival in AMD patients following choroidal new vessel (CNV) excision.

描述（由申请人提供）：已提议用健康的视网膜色素上皮（RPE）重新铺面，或者对生病的BRUCH膜（RPE）的膜，以作为与年龄相关的黄斑变性（AMD）的辅助治疗。但是，老化的下Bruch膜不支持移植的RPE存活和分化。随着衰老和AMD的衰老，Bruch的膜经历了许多可能影响配体可用性的变化。这些观察结果导致了以下假设：衰老会改变掩盖细胞外基质配体的可用性，以移植后的RPE细胞，损害了连接后事件，并导致细胞死亡或细胞无法区分的细胞死亡或无能为力。以下特定目的将通过用牛角膜内皮细胞外基质（BCE-ECM）重新铺面，该基质是支持RPE生长和分化的基质，将确定是否可以改善细胞健康。目标1：确定牛角膜内皮细胞（BCE）是否可以“重新表面” Bruch的膜。 AIM 2：确定培养的RPE细胞是否可以在BCE-ECM涂层的RPE基底膜/BRUCH膜上形成焦点粘连，以及是否改善了短期细胞存活。 AIM 3：确定RPE细胞在BCE-ECM涂层的BRUCH膜上是否改善了长期生存。这些研究将利用人类下Bruch膜的器官外植体培养系统。重新铺面将通过在Epplants上培养BCE一段时间来实现，以允许BCE-ECM沉积，然后去除细胞并用培养的RPE细胞播种。外植体将通过扫描，光和共聚焦显微镜进行分析。如果通过这种方法增强了RPE的存活率和分化，那么研究将为进一步的研究奠定基础，以使用BCE-ECM的配体成分进行进一步的研究，这可能为BRUCH膜的临床治疗构成基础，以增强脉络膜新患者的RPE移植物存活率血管（CNV）切除。