CULTURED CILIATED CELLS FOR HIGH THROUGHPUT SCREENING

描述(申请人提供)：BioTechPlex计划开发哺乳动物气管/支气管呼吸道上皮细胞系CILSCREEN TM，专门用于高通量筛查。这是基于生理机能分析。BioTechPlex已经完成了所有提出的第一阶段目标。A)成功开发了培养和数值扩增呼吸道上皮细胞的技术，同时保留了纤毛发生和纤毛功能。B)开发了重新启动纤毛发生机制以在浸泡培养导致纤毛丧失后再生纤毛的方法。C)开发了诱导细胞极化的方法，以诱导超过50%的上皮细胞上纤毛生长，这些上皮细胞至少保持正常纤毛跳动3-4周。在这个第二阶段的项目中，BioTechPlex将结合和扩展第一阶段的成果，生产五个物种的可行商业细胞系，形成第一个哺乳动物纤毛细胞库。具体地说，BioTechPlex将实现以下目标。1)。进一步确定调控细胞分裂和分化的因素。2)。产生大量的呼吸道上皮纤毛细胞。3)。使用生物标志物、形态特征和生理功能来表征细胞系；建立哺乳动物纤毛细胞库。BioTechPlex计划促进呼吸道纤毛上皮细胞在细胞功能研究中的广泛使用，即这些细胞将成为用于高通量筛选肺部药物发现的成纤维细胞、肿瘤细胞系和其他基因改变细胞系的替代品。这些细胞模型体系将适用于正常和诱导病理机制及其相关药理学的研究。

DESCRIPTION (provided by applicant): BioTechPlex proposes to develop mammalian tracheal/bronchial airway epithelial cell line, CILSCREEN TM, specifically for high throughput screening. This is based on physiological functional assays. BioTechPlex has accomplished all the proposed Phase I objectives. A) Successfully developed techniques to culture and numerically expand airway epithelial cells while preserving ciliogenesis and ciliary function. B) Developed methodologies to reactivate mechanisms of ciliogenesis to regenerate the cilia following loss of cilia induced by submerged culture. C) Developed methodologies to induce cell polarization so as to induce cilia outgrowth on more than 50 percent of epithelial cells, which maintained normal ciliary beating for at least 3-4 weeks. In this Phase II project, BioTechPlex will incorporate and extend the accomplishments of Phase I to produce viable commercial cell lines in five species to form the first Mammalian Ciliated Cell Bank. Specifically, BioTechPlex will accomplish the following objectives. 1). Further identify factors regulating cell division and differentiation. 2). Produce large quantities of airway epithelial ciliated cells. 3). Characterize the cell lines using biomarkers, morphological characteristics and physiological function, and 4). Establish a Mammalian Ciliated Cell Bank. BioTechPlex plans to facilitate widespread use of airway ciliated epithelial cells for cell function studies, i.e. the cells will become an alternative to the use of fibroblasts, tumor cell lines and other genetically altered cell lines utilized in high throughput screening for pulmonary drug discovery. These cell model systems will be suitable for the study of normal and induced pathological mechanisms and their related pharmacology.