Expression of human dystrophin following the transplantation of genetically modified mdx myoblasts

Abstract

Transplantation of genetically modified autologous myoof SCID mice. Ten and 24 days after transplantation, blasts has been proposed as a possible solution to avoid numerous muscle fibers expressing both human dystrophin long-term use of immunosuppressive drugs. To determine and ␤-Gal were detected throughout the mouse muscles the conditions to be used in this kind of approach for possby immunohistochemistry using an antibody specific for ible treatment of dystrophin deficiency, mdx myoblasts human dystrophin. The presence of the human mini-dystrowere infected at different multiplicities of infection (MOI of phin mRNA was also detected by RT-PCR. These results 0.01-1000) with an adenoviral vector containing a CMV demonstrate that three essential conditions in autologous promoter/enhancer driven 6.3 kb human dystrophin cDNA myoblast transplantation can be achieved: (1) in vivo sur-(minigene) and tested in vitro for transgene expression. In vival of at least some of the transduced myoblasts; (2) these cultures, dystrophin mRNA was found to be proefficient fusion of these cells with the host muscle fibers; portionate with increasing MOI. Primary myoblast cultures and (3) the high expression of the dystrophin transgene in derived from transgenic mdx mice expressing ␤-Gal under situ. Furthermore, this article provides a novel RT-PCRa muscle-specific promoter and showing high expression based technique to quantify the human dystrophin miniof the human mini-dystrophin transgene introduced by the gene expression in vitro and in vivo. adenoviral vector were grafted into anterior tibialis muscles

Stable transduction of myogenic cells with lentiviral vectors expressing a minidystrophin

“…This type of immune response was not eliminated by using lentiviral vectors bearing a muscle-specific promoter (data not shown), consistent with observations by others using either adenoviral (Ad) or herpes viral vectors for autologous transplantation of mdx mice. [20][21][22] Similarly, mdx MAPCs genetically modified with microdystrophin regulated by a musclespecific promoter, when injected intramuscularly, also generated dystrophin-expressing myofibers in mdx skeletal muscles (Reyes et al, submitted).…”

Cell Transplantation and “Stem Cell Therapy” in the Treatment of Myopathies: Many Promises in Mice, Few Realities in Humans

“…This strategy avoids incompatibility of the Major Histocompatibility Complex among the recipient and the transplanted mononuclear cells, but leaves the possibility of eliciting a specific immune response if the genetically-modified cells express epitopes not shared with the recipient [199]. Since the first experiments that used adenoviral vectors [200][201][202], the range of possibilities of genetic manipulation of the cells has been growing. Among viral vectors, lentiviruses attract much attention because they can infect postmitotic or quiescent cells and integrate the transgene in the host genome, providing stable long-term gene expression both in vitro and in vivo [203,204].…”

Chondrogenic differentiation of canine myoblasts induced by cartilage-derived morphogenetic protein-2 and transforming growth factor-β1 in vitro

“…Myoblasts are regarded as ideal donor cells due to their abundant source, obtainability, stem cell-like potential, extended survival ability and rapid proliferative activity in vitro (8). Myoblasts are considered superior in biopsy, manipulation and clinical application than other stem cells, such as chondrocytes or bone mesenchymal stem cells (9). Therefore, myoblasts may be suitable for providing autogenous cells for the treatment of meniscal injuries.…”