Co-Transplantation of Plasmid-Transfected Myoblasts and Myotubes into Rat Brains Enables High Levels of Gene Expression Long-Term
We have previously proposed the use of primary muscle cells as a “platform,” or “vehicle” for intracerebral transgene expression. Brain grafts of minced muscle, or cultured muscle cells persisted in rat brains for at least 6 mo without any decrease in graft size, or tumor formation. Stable, but mode...
Main Authors: | , , , , |
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Format: | Article |
Language: | English |
Published: |
SAGE Publishing
1993-05-01
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Series: | Cell Transplantation |
Online Access: | https://doi.org/10.1177/096368979300200302 |
Summary: | We have previously proposed the use of primary muscle cells as a “platform,” or “vehicle” for intracerebral transgene expression. Brain grafts of minced muscle, or cultured muscle cells persisted in rat brains for at least 6 mo without any decrease in graft size, or tumor formation. Stable, but moderate levels of intracerebral transgene expression were obtained by transplanting plasmid-transfected myotubes in culture. In the present study, high and stable levels of intracerebral transgene expression were achieved by the co-transplantation of plasmid-transfected myoblasts and myotubes in culture. Approximately 5 × 10s myoblasts and myotubes were transfected with 10 μg pRSVL plasmid DNA, and 30 μg Lipofectin (BRL), respectively. They were mixed together (total cell number was 1 million), and stereotactically injected into the caudate nucleus of an adult rat brain. The activity of luciferase, the product of transgene expression, was stable for at least 4 mo, and much higher than the levels in myotube grafts, or co-grafts of myoblasts and minced muscle. Presumably, the myotubes served as a framework on which the myoblasts can form myotubes. The sections of brains transplanted with co-graft of myoblasts, and myotubes transfected with pRSVLac-Z were stained immunofluorescently for β-galactosidase activity. The muscle grafts contained β-galactosidase positive myofibers 4 mo after transplantation. Such high and stable levels of in vivo expression after postnatal gene transfer have rarely been achieved. Primary muscle cells are useful vehicle for transgene expression in brains, and potentially valuable for gene therapy of degenerative neurological disorders. |
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ISSN: | 0963-6897 1555-3892 |