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.