GENE TRANSFER INTO LIVER AND HEMAPOIETIC/THYMIC STEM CELLS

基因转移至肝脏和造血/胸腺干细胞

• 批准号: 3878954
• 负责人: L BALTRUCKI
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3878954
• 关键词: CD4 molecule; gene expression; gene therapy; genetic manipulation; genetic models; hematopoietic stem cells; hematopoietic tissue; liver cells; liver regeneration; mixed tissue /cell culture; model design /development; molecular genetics; thymus; transposon /insertion element

Retroviral vectors have been developed as an efficient system for delivering genes into many types of eukaryotic cells, in vitro. Success with in vivo gene transfer, however, has been limited by the lack of an effective delivery system. To address this problem, we are; 1. developing methods for introducing vectors or transduced hepatocytes/stem cells, directly into regenerating liver, initially in the rat animal model, and 2. attempting to create implantable bioartificial devices, which will function as substitutes for liver tissue, capable of expressing genes with therapeutic potential, indefinitely. The efficiency of gene transfer into the self-renewing hematopoietic/lymphoid stem cells is too low for retroviralmediated gene transfer to be applied successfully to human gene therapy. In an effort to improve the efficiency of gene transfer into stem cells, a three-dimensional culture system, based on a hollow-fiber/microcarrier perfusion bioreactor, is under development. In this system, bone marrow stromal cells or thymic epithelial cells will be cultured with the CD34+ marrow cell population, to high cell densities. These co-cultures will serve as "affinity columns" for pluripotent and T-lymphoid stem cells respectively.

逆转录病毒载体已被开发为一种有效的系统 在体外将基因导入多种类型的真核细胞。成功 然而，体内基因转移一直受到缺乏 有效的交付系统。为了解决这个问题，我们正在：1.发展 导入载体或转导的肝细胞/干细胞的方法， 直接进入再生肝，最初在大鼠动物模型中，和2。 试图创造可植入的生物人工设备，它将发挥作用 作为肝组织的替代品，能够表达基因 无限期的治疗潜力。 基因转移在自我更新中的效率 造血/淋巴干细胞对于逆转录病毒介导的基因来说太低了 转移成功应用于人类基因治疗。为了努力 提高干细胞基因转移的效率，a 基于中空纤维/微载体的三维培养系统 灌流生物反应器正在开发中。在这个系统中，骨髓 基质细胞或胸腺上皮细胞将与CD34+一起培养 骨髓细胞数量，到高细胞密度。这些共同文化将会 充当多能干细胞和T淋巴细胞的“亲和柱” 分别进行了分析。