학술논문
Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice.
Document Type
Academic Journal
Author
O'Neill A; Department of Physiology, University of Maryland School of Medicine, Baltimore, MD, USA.; Martinez AL; Department of Physiology, University of Maryland School of Medicine, Baltimore, MD, USA.; Mueller AL; Department of Physiology, University of Maryland School of Medicine, Baltimore, MD, USA.; Cell Metabolism, Cambridge, MA, USA.; Huang W; Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD, USA.; Accorsi A; Fulcrum Therapeutics, Cambridge, MA, USA.; Blackbird Laboratories, Baltimore, MD, USA.; Kane MA; Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD, USA.; Eyerman D; Fulcrum Therapeutics, Cambridge, MA, USA.; Apellis Pharmaceuticals, Waltham, MA, USA.; Bloch RJ; Department of Physiology, University of Maryland School of Medicine, Baltimore, MD, USA.
Source
Publisher: SAGE Publishing Country of Publication: United States NLM ID: 9208854 Publication Model: Print Cited Medium: Internet ISSN: 1555-3892 (Electronic) Linking ISSN: 09636897 NLM ISO Abbreviation: Cell Transplant Subsets: MEDLINE
Subject
Language
English
Abstract
Xenografts of human skeletal muscle generated in mice can be used to study muscle pathology and to test drugs designed to treat myopathies and muscular dystrophies for their efficacy and specificity in human tissue. We previously developed methods to generate mature human skeletal muscles in immunocompromised mice starting with human myogenic precursor cells (hMPCs) from healthy individuals and individuals with facioscapulohumeral muscular dystrophy (FSHD). Here, we examine a series of alternative treatments at each stage in order to optimize engraftment. We show that (i) X-irradiation at 25Gy is optimal in preventing regeneration of murine muscle while supporting robust engraftment and the formation of human fibers without significant murine contamination; (ii) hMPC lines differ in their capacity to engraft; (iii) some hMPC lines yield grafts that respond better to intermittent neuromuscular electrical stimulation (iNMES) than others; (iv) some lines engraft better in male than in female mice; (v) coinjection of hMPCs with laminin, gelatin, Matrigel, or Growdex does not improve engraftment; (vi) BaCl 2 is an acceptable replacement for cardiotoxin, but other snake venom preparations and toxins, including the major component of cardiotoxin, cytotoxin 5, are not; and (vii) generating grafts in both hindlimbs followed by iNMES of each limb yields more robust grafts than housing mice in cages with running wheels. Our results suggest that replacing cardiotoxin with BaCl 2 and engrafting both tibialis anterior muscles generates robust grafts of adult human muscle tissue in mice.
Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.