■开发适用于疾病状况和治疗的体外研究的3维角膜构建体。
■使用化学交联的胶原蛋白和硫酸软骨素细胞外基质创建体外人角膜构建体,并接种3种人角膜细胞类型(上皮,基质,和内皮细胞)以及神经细胞。神经细胞来自杂合神经母细胞瘤细胞,其他细胞来自永生化人角膜细胞系。为了检查可行性并描述结构的特征,细胞毒性,细胞增殖,组织学,和蛋白质表达研究。
■优化的培养条件允许跨构建体内的细胞类型的同步生存力。该结构显示了不同细胞层的典型外观,包括健康的外表,表型分化的神经元。用蛋白质印迹确认构建体中特定细胞类型的预期蛋白质表达谱。
■成功开发了一种体外角膜构建体,并维持了具有解剖学上正确的细胞基因座的单个细胞表型。该构建体可用于评估特定的角膜疾病和开发不同的角膜疾病模型。此外,所述构建体可用于评价药物靶向和/或对单个角膜层的渗透,测试角膜疾病的新疗法,并潜在地降低了在研究的早期阶段对动物进行角膜研究的必要性。
■专有或商业披露可在本文末尾的脚注和披露中找到。
UNASSIGNED: To develop a 3-dimensional corneal construct suitable for in vitro studies of disease conditions and therapies.
UNASSIGNED: In vitro human corneal constructs were created using chemically crosslinked collagen and chondroitin sulfate extracellular matrix and seeded with 3 human corneal cell types (epithelial, stromal, and endothelial) together with neural cells. The neural cells were derived from hybrid neuroblastoma cells and the other cells used from immortalized human corneal cell lines. To check the feasibility and characterize the constructs, cytotoxicity, cell proliferation, histology, and protein expression studies were performed.
UNASSIGNED: Optimized culture condition permitted synchronized viability across the cell types within the construct. The construct showed a typical appearance for different cellular layers, including healthy appearing, phenotypically differentiated neurons. The expected protein expression profiles for specific cell types within the construct were confirmed with western blotting.
UNASSIGNED: An in vitro corneal construct was successfully developed with maintenance of individual cell phenotypes with anatomically correct cellular loci. The construct may be useful in evaluation of specific corneal disorders and in developing different corneal disease models. Additionally, the construct can be used in evaluating drug targeting and/or penetration to individual corneal layers, testing novel therapeutics for corneal diseases, and potentially reducing the necessity for animals in corneal research at the early stages of investigation.
UNASSIGNED: Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.