Core B--Tissue Procurement and Cell Culture Core

核心B--组织获取和细胞培养核心

• 批准号: 6956040
• 负责人: Scott H Randell
• 金额: $ 10.19万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-20 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6956040
• 关键词: biological transport; biomedical facility; bronchial mucus; cryopreservation; cystic fibrosis; gene therapy; human tissue; immunocytochemistry; in situ hybridization; respiratory epithelium; tissue /cell culture; tissue /cell preparation; tissue resource /registry

Cell cultures are instrumental for studying basic and applied aspects of gene therapy for cytic fibrosis (CF). Human airway epithelial cells maintained at an air-liquid interface display mucociliary differentiation similar to the in vivo epithelium. These cells faithfully reproduce the high resistance to vectors, characteristic of the native mucosa, providing a strong platform for pre-clinical studies to address issues crucial to the success of gene therapy for CF. A Tissue Procurement and Cell Culture Core was established at the University of North Carolina (UNC) in 1984, under the auspices of the CF Foundation, to provide standardized cell cultures to CF researchers. The Core has supported UNC Gene Therapy for CF projects since 1993 and has increased its output and capabilities to meet growing research demands. The Core routinely makes available cells and media that are unavailable and/or prohibitively expensive if purchased commercially. The present application will provide continuity in the supply of human airway epithelial cell cultures to UNC Gene Therapy for Cystic Fibrosis Program Project investigators by providing the following specific functions: 1) Obtain airway tissues from normal, CF and disease control humans as the source for primary cells, for acute ex vivo experiments, protein and RNA isolation, immunostaining and in situ hybridization. Characterize donors while protecting confidentiality. 2) Isolate primary epithelial cells from excised human airways. Prepare and maintain primary epithelial cell cultures. Provide support for preparation of well differentiated airway epithelial cultures from passaged or cryopreserved primary human cells. Optimize culture conditions to replicate gene expression, morphological differentiation, and physiologic functions of airways as evidenced by rotational mucus transport and regulation of the airway surface liquid (ASL) layer. Through these functions, and in conjunction with other Cores in this application, the Cell Culture Core will support research towards the development of safe and effective gene therapy for CF.

细胞培养对于研究细胞纤维化（CF）的基本和应用方面具有重要作用。维持在空气界面上的人类气道上皮细胞显示的粘膜钙化分化与体内上皮相似。这些细胞忠实地再现了对载体的高抗性，这是天然粘膜的特征，为临床前研究提供了一个强大的平台，以解决对基因治疗对CF的成功至关重要的问题。 1984年，在CF基金会的主持下，在北卡罗来纳大学（UNC）建立了组织采购和细胞培养核心，为CF研究人员提供了标准化的细胞培养。该核心自1993年以来一直支持CF项目的UNC基因疗法，并提高了其产量和能力，以满足不断增长的研究需求。核心通常使可用的单元和媒体在商业上购买，这些细胞和媒体不可用和/或过于昂贵。本应用将在人类气道上皮细胞培养物的供应中提供连续性，以提供囊性纤维化计划项目研究人员的基因疗法。 特定功能：1）从正常，CF和疾病控制人类作为原代细胞的来源中获得气道组织，用于急性外体实验，蛋白质和RNA分离，免疫染色和原位杂交。在保护机密性的同时，以捐助者为特征。 2）与切除的人类气道分离原代上皮细胞。准备并维持原发性上皮细胞培养物。为从传统或冷冻保存原代人细胞制备良好分化的气道上皮培养物提供了支持。优化培养条件以复制气道的基因表达，形态分化和生理功能，如旋转粘液传输和气道表面液体（ASL）层的调节所证明的那样。通过这些功能，并与该应用中的其他内核结合在一起，该单元格 培养核心将支持为CF开发安全有效的基因疗法的研究。