The Effect of Growth Facotrs and Extracellular Matrix Materials on the Growth and Differentiation of Microencapsulated Myoblasts

Abstract

An alternative gene therapy method, non-autologous somatic-gene therapy, is the use of a genetically modified universal cultured cell line that can be implanted into different allogeneic recipients. When used as recombinant cells in microcapsules, myoblasts possess several advantages over other cell types, namely their ability to terminally differentiate thus preventing overcrowding within the capsular space. However, encapsulated myoblasts demonstrate decreased proliferation and myogenic differentiation when compared to unencapsulated myoblasts due to the unnatural capsule environment. This study aims to improve the microcapsule environment by incorporating basic fibroblast growth factor (bFGF) and insulin-like growth factor-11 (IGF-11) and the extracellular matrix materials, collagen, laminin-1 and merosin (laminin-2) within the microcapsules in an attempt to mimic the natural surrounding required for myoblast growth and differentiation. While bFGF lead to significant increases in encapsulated myoblast proliferation, it did not appear to be an ideal choice for optimizing the microcapsule environment due to its inhibitory effect on differentiation and the relative cost in therapeutic delivery of proteins. Both merosin and the combination of laminin and merosin together provide a better alternative for increasing myoblast growth and survival within microcapsules since they have no apparent inhibitory effect on myogenic differentiation, and produce similar proliferative results seen when using bFGF. In terms of differentiation, the addition of IGF-11 to the microcapsules or the use of a myoblast cell line overexpressing IGF-11, aid in increasing the myogenic differentiation of encapsulated myoblasts, however, differentiation levels still do not approach those seen in unencapsulated myoblasts. The positive results obtained with the growth factors and matrix materials employed in this study are important steps towards the optimization of microcapsules by improving both the proliferation and differentiation of encapsulated myoblasts. However, more study is needed to elucidate possible solutions to the continued problem of decreased differentiation of myoblasts within APA microcapsules in order to achieve myogenic differentiation that is comparable to what is seen in unencapsulated myoblasts.