Acoustically-Enhanced Myogenic Cell Culture

声学增强的生肌细胞培养

• 批准号: 6736983
• 负责人: TODD A MCADAMS
• 金额: $ 11.92万
• 依托单位: RESODYN CORPORATION
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-18 至 2007-02-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6736983
• 关键词: bioengineering /biomedical engineering; biomedical equipment development; bioreactors; biotechnology; carbon dioxide; cell proliferation; growth media; mass tissue /cell culture; mechanical stress; muscle cells; oxygen consumption; sound frequency; technology /technique development

DESCRIPTION (provided by applicant): The use of human muscle cells for tissue engineering applications holds significant promise for new treatments of muscle-cell disorders, organ repair, and gene therapy. With the enabling advances in biology and immunology nearing fruition, the next challenge to overcome is the simple and economical production of large numbers (approximately 10/11 cells) of adherent-dependent muscle cells. The problem is succinctly summarized by one of the leading research groups in the muscle-cell transplantation field: "The major problem encountered in myoblast culture scale-up is that these primary, anchorage-dependent cells are known to be particularly sensitive to hydrodynamic stresses and to grow very slowly, if at all, on surfaces exposed to even minute fluid movement." Resodyn Corporation intends to develop novel tissue culture methods to enable the efficient production of shear-sensitive primary adherent cell types for cellular therapy applications. The specific aim of this Phase I proposal is to demonstrate enhanced myogenic cell culture productivity (compared to conventional techniques) through the use of acoustically-agitated microcarrier culture. The potential therapeutic applications of muscle cells and muscle-derived stem ceils are broad, and include treatment of muscular dystrophies, use as a vehicle for gene therapy protocols, repair of damaged heart tissue, incontinence repair, artificial blood vessels, and repair of bone and cartilage defects. These novel tissue culture methods will be based on the use of ResonantSonics(c)(RS), a new method of mixing that utilizes propagation of low-frequency acoustic energy waves through a liquid medium. While providing superior mass transfer (increased oxygen transfer and reduced mixing times) to standard impeller- or spinner-based systems, RS dramatically reduces the amount of fluid shear stress experienced by cells and particles in the liquid medium. It is therefore proposed to utilize RS as the basis for a highly productive acoustically mixed cell culture system for the large scale production of human myogenic cells. In order to achieve this goal, the proposed research will examine the proper acoustic intensity, microcarrier type, and microcarrier density necessary to achieve maximal growth of primary myogenic cells. An optimized acoustic culture system will be compared to current state-of-the-art protocols to demonstrate superior efficacy. Successful feasibility demonstration of this technique will have substantial additional utility for the production of other types of primary and immortalized adherent cells.

描述（由申请人提供）：使用人肌细胞进行组织工程应用，对于肌细胞疾病、器官修复和基因治疗的新疗法具有重要的前景。 随着生物学和免疫学的进展接近成熟，下一个需要克服的挑战是简单而经济地生产大量（约10/11细胞）的粘附依赖性肌细胞。肌肉细胞移植领域的一个领先研究小组简洁地总结了这个问题： “成肌细胞培养规模扩大遇到的主要问题是，这些主要的锚定依赖性细胞已知对流体动力学应力特别敏感，并且在暴露于即使是微小的流体运动的表面上生长非常缓慢。" Resodyn公司打算开发新的组织培养方法，使细胞治疗应用的剪切敏感性原代贴壁细胞类型的有效生产。该I期提案的具体目的是通过使用声搅拌微载体培养来证明增强的肌原性细胞培养生产率（与常规技术相比）。肌细胞和肌源性干细胞的潜在治疗应用是广泛的，并且包括肌营养不良的治疗、用作基因治疗方案的载体、受损心脏组织的修复、失禁修复、人工血管以及骨和软骨缺陷的修复。这些新型组织培养方法将基于ResonantSonics（c）（RS）的使用，这是一种利用低频声能波通过液体介质传播的新混合方法。在提供优于标准叶轮或基于旋转器的系统的上级传质（增加的氧传递和减少的混合时间）的同时，RS显著地减少了液体培养基中的细胞和颗粒所经历的流体剪切应力的量。因此，提出利用RS作为用于大规模生产人肌原性细胞的高产声混合细胞培养系统的基础。为了实现这一目标，拟议的研究将检查适当的声强度，微载体类型和微载体密度，以实现原代肌源性细胞的最大生长。将优化的声培养系统与当前最先进的方案进行比较，以证明其上级功效。该技术的成功可行性证明将对其他类型的原代和永生化贴壁细胞的生产具有实质性的额外效用。