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）：将人类肌肉细胞用于组织工程应用，对肌肉细胞疾病，器官修复和基因疗法的新疗法具有重要的希望。 随着生物学和免疫学取得的促进，要克服的下一个挑战是，依赖性肌肉细胞的大量（约10/11个细胞）的简单而经济地生产。肌肉细胞移植领域的主要研究小组之一简洁地总结了这个问题： “在成肌细胞培养中遇到的主要问题是，已知这些主要的锚固依赖性细胞对流体动力胁迫特别敏感，并且在暴露于均匀的流体运动的表面上非常缓慢地生长。” Resodyn Corporation打算开发新型的组织培养方法，以使剪切敏感的原发性粘附细胞类型有效地生产用于细胞治疗的应用。该阶段I提出的具体目的是通过使用声学的微载体培养来证明增强的肌源性细胞培养生产力（与常规技术相比）。肌肉细胞和肌肉衍生的茎天花板的潜在治疗应用是广泛的，包括治疗肌肉营养不良，用作基因治疗方案的载体，修复受损的心脏组织，失禁修复，人造血管以及骨骼和软骨缺陷的修复。这些新型的组织培养方法将基于使用resonantonics（C）（RS）的使用，这是一种新的混合方法，该方法利用低频声能通过液体培养基传播。在为标准的叶轮或基于旋转器的系统提供上质传质（增加氧气转移和减少混合时间）的同时，RS大大减少了液体培养基中细胞和颗粒所经历的流体剪切应力的量。因此，提议将RS用作人类肌原性细胞大规模生产的高生产性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。将优化的声学培养系统与当前的最新协议进行比较，以证明效率较高。成功的可行性证明了该技术将具有大量额外的效用，以生产其他类型的原发性和永生化的粘附细胞。