Automated Large-Scale Production of Endothelial Cells

内皮细胞的自动化大规模生产

• 批准号: 6693921
• 负责人: Kristin Goltry
• 金额: $ 9.98万
• 依托单位: AASTROM BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2004-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6693921
• 关键词: bioengineering /biomedical engineering; biomedical equipment development; bioreactors; biotechnology; bone marrow; cell population study; clinical research; fibronectins; growth media; human tissue; laboratory mouse; stem cells; tissue /cell culture; tissue /cell preparation; vascular endothelium

DESCRIPTION (provided by applicant): Treatment of ischemia by administration of bone marrow (BM)-derived endothelial progenitor cells (EPC) or ex vivo expanded EPC populations has been shown to enhance vascular regeneration in animal models and in clinical trials. For cell therapy to be practical in a clinical setting, simple, efficient, GMP compliant methods for cell production are needed. Aastrom has previously developed a single-pass perfusion (SPP) bioreactor technology, the AastromReplicell TM Cell Production System (ARS), designed for automated expansion of primary human cells for clinical use. In previous studies, SPP technology has been shown to enhance function and overall production of several cell types. Based on this technology, we will develop a GMP I compliant method for the ex vivo production of substantial numbers of EPC from a small volume of BM for use in cell therapy applications. First, we will identify and optimize EPC cell culture parameters in our small-scale SPP culture system. Parameters to be examined will include starting cell population, medium serum concentration, culture duration, starting cell density and frequency of medium exchange. EPC will be enumerated and output analyzed by immunostaining and in vitro assays. Optimal culture conditions from Aim 1 will be evaluated in an in vivo model for hind limb ischemia in Aim 2. Concurrently, we will begin studies in Aim 3 to develop the EPC culture process(es) in the clinical-scale ARS. Based on small-scale results, potential revisions to the current ARS platform technology will include cell bed surface modifications and software updates to accommodate optimized medium perfusion rates. These modifications will be made with the help of in-house engineers and product specialists. Cell output from the ARS will be compared to cell output from small-scale cultures. With the successful completion of Phase I studies, a Phase II program will involve continued culture optimization and in vivo evaluations of the expanded cell products in both small-scale and clinical-scale cultures to optimize the ARS for large-scale clinical trial commercialization. Once the ARS process is established, a clinical trial will be initiated for the treatment of vascular disease. Studies will also be initiated to develop EPC gene transfer protocols within the context of the ARS platform. This work will result in the manufacture of a cell therapy kit offering automated GMP production of functional EPC cells for use in treatment of peripheral and cardiovascular diseases.

描述（由申请人提供）：在动物模型和临床试验中，通过给予骨髓（BM）衍生的内皮祖细胞（EPC）或离体扩增的EPC群治疗缺血已显示出增强血管再生。为了使细胞疗法在临床环境中实用，需要简单、有效、符合GMP的细胞生产方法。Aastrom先前开发了一种单程灌注（SPP）生物反应器技术，即AastromReplicell TM细胞生产系统（ARS），旨在自动扩增临床使用的原代人类细胞。在以前的研究中，SPP技术已被证明可以增强几种细胞类型的功能和整体生产。基于该技术，我们将开发一种符合GMP I的方法，用于从小体积BM中离体生产大量EPC，用于细胞治疗应用。首先，我们将在我们的小规模SPP培养系统中确定和优化EPC细胞培养参数。待检查的参数包括起始细胞群、培养基血清浓度、培养持续时间、起始细胞密度和培养基更换频率。将通过免疫染色和体外试验对EPC进行计数和输出分析。将在目标2的后肢缺血体内模型中评价目标1的最佳培养条件。同时，我们将开始目标3中的研究，以开发临床规模ARS中的EPC培养工艺。根据小规模结果，对当前ARS平台技术的潜在修改将包括细胞床表面修改和软件更新，以适应优化的培养基灌注速率。这些修改将在内部工程师和产品专家的帮助下进行。将ARS的细胞输出与小规模培养的细胞输出进行比较。随着I期研究的成功完成，II期计划将涉及持续的培养优化和在小规模和临床规模培养中对扩增细胞产品的体内评价，以优化ARS用于大规模临床试验商业化。一旦建立了ARS过程，将启动血管疾病治疗的临床试验。还将启动研究，以在ARS平台范围内制定EPC基因转移方案。这项工作将导致细胞治疗试剂盒的生产，提供用于治疗外周和心血管疾病的功能性EPC细胞的自动GMP生产。