Development of Approaches to Improve Cell Survival in Myoblast Transfer Therapy

Abstract

Myoblast transplantation has been extensively studied as a gene complementation approach for genetic diseases such as Duchenne Muscular Dystrophy. This approach has been found capable of delivering dystrophin, the product missing in Duchenne Muscular Dystrophy muscle, and leading to an increase of strength in the dystrophic muscle. This approach, however, has been hindered by numerous limitations, including immunological problems, and low spread and poor survival of the injected myoblasts. We have investigated whether antiinflammatory treatment and use of different populations of skeletal muscle–derived cells may circumvent the poor survival of the injected myoblasts after implantation. We have observed that different populations of muscle-derived cells can be isolated from skeletal muscle based on their desmin immunoreactivity and differentiation capacity. Moreover, these cells acted differently when injected into muscle: 95% of the injected cells in some populations died within 48 h, while others richer in desmin-positive cells survived entirely. Since pure myoblasts obtained from isolated myofibers and myoblast cell lines also displayed a poor survival rate of the injected cells, we have concluded that the differential survival of the populations of muscle-derived cells is not only attributable to their content in desmin-positive cells. We have observed that the origin of the myogenic cells may influence their survival in the injected muscle. Finally, we have observed that myoblasts genetically engineered to express an inhibitor of the inflammatory cytokine, IL-1, can improve the survival rate of the injected myoblasts. Our results suggest that selection of specific muscle-derived cell populations or the control of inflammation can be used as an approach to improve cell survival after both myoblast transplantation and the myoblast-mediated ex vivo gene transfer approach.

Over-expression of CXCR4 on mesenchymal stem cells augments myoangiogenesis in the infarcted myocardium

“…Although MSCs have been shown to participate in myocardial repair, their homing to infarcted area and their survival post transplantation are limited [16]. The cell viability may ultimately determine the final outcome of cardiac repair [17]. CXCR4 is receptor for SDF-1α and is very important in the stem cell migration, homing, and survival [18].…”

Muscle-derived cell-mediated ex vivo gene therapy for urological dysfunction

“…Previous experiments have shown that early preplates (pp1-2) contain a majority of fibroblasts (5-15% desmin-positive) and the late preplates (pp5-6) are enriched with desmin-positive cells (>80%). [20][21][22]41 The cells used in this experiment were taken from the late preplate (pp6) and were shown to express myogenic and stem cell markers. 9 The proliferation medium used to grow the cells was Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 10% horse serum (HS) and 1% penicillin/streptomycin.…”

“…[7][8][9][10] However, the transplantation of donor muscle cells into host muscle has been hindered by various limitations, such as immune rejection that consequently leads to poor spreading and survival of injected cells. [11][12][13][14][15][16][17][18][19][20] The success of cell transplantation may be dramatically improved by using a specific population of cells that has the capacity to overcome these hurdles.…”

“…20 These cells were harvested from a muscle biopsy and purified by the preplate technique based on their adherence characteristics to collagen-coated flasks. [20][21][22] The purified MDC (pp6) used for these experiments took about 6 days to adhere to collagen-coated flasks and stained positive for myogenic (desmin) and well-known stem cell markers, including Sca-1 and FLK-1. 9 These results suggest that this population of highly purified MDC may contain stem cells.…”

Stem Cell Monitoring with a Direct or Indirect Labeling Method

“…Depending on the imaging modality to be used, stem cells can be labeled with radionuclides for SPECT or PET [32][33][34][35] and SPIO for MRI (Fig. 1) [36,37]. Direct cell labeling is fairly simple and does not involve genetic modification of the cell.…”