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Mol Ther Methods Clin Dev. 2014 Sep 10;1:14040. doi: 10.1038/mtm.2014.40. eCollection 2014.

Developmental stage determines efficiency of gene transfer to muscle satellite cells by in utero delivery of adeno-associated virus vector serotype 2/9.

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1
The Children's Center for Fetal Research, Children's Hospital of Philadelphia , Philadelphia, Pennsylvania, USA ; Department of Pediatric Surgery, Yale School of Medicine , New Haven, Connecticut, USA.
2
The Children's Center for Fetal Research, Children's Hospital of Philadelphia , Philadelphia, Pennsylvania, USA.
3
Department of Pathology and Laboratory Medicine, Gene Therapy Program, Perelman School of Medicine at the University of Pennsylvania , Philadelphia, Pennsylvania, USA.

Abstract

Efficient gene transfer to muscle stem cells (satellite cells) has not been achieved despite broad transduction of skeletal muscle by systemically administered adeno-associated virus serotype 2/9 (AAV-9) in mice. We hypothesized that cellular migration during fetal development would make satellite cells accessible for gene transfer following in utero intravascular injection. We injected AAV-9 encoding green fluorescent protein (GFP) marker gene into the vascular space of mice ranging in ages from post-coital day 12 (E12) to postnatal day 1 (P1). Satellite cell transduction was examined using: immunohistochemistry and confocal microscopy, satellite cell migration assay, myofiber isolation and FACS analysis. GFP positive myofibers were detected in all mature skeletal muscle groups and up to 100% of the myofibers were transduced. We saw gestational variation in cardiac and skeletal muscle expression. E16 injection resulted in 27.7 ± 10.0% expression in satellite cells, which coincides with the timing of satellite cell migration, and poor satellite cell expression before and after satellite cell migration (E12 and P1). Our results demonstrate that efficient gene expression is achieved in differentiated myofibers and satellite cells after injection of AAV-9 in utero. These findings support the potential of prenatal gene transfer for muscle based treatment strategies.

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