Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice
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 immunoc...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2024-04-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.1177/09636897241242624 |
| _version_ | 1797213915553202176 |
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| author | Andrea O’Neill Anna Llach Martinez Amber L. Mueller Weiliang Huang Anthony Accorsi Maureen A. Kane David Eyerman Robert J. Bloch |
| author_facet | Andrea O’Neill Anna Llach Martinez Amber L. Mueller Weiliang Huang Anthony Accorsi Maureen A. Kane David Eyerman Robert J. Bloch |
| author_sort | Andrea O’Neill |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-24T11:05:52Z |
| format | Article |
| id | doaj.art-f06ef38857ad477f832efdf881641864 |
| institution | Directory Open Access Journal |
| issn | 1555-3892 |
| language | English |
| last_indexed | 2024-04-24T11:05:52Z |
| publishDate | 2024-04-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-f06ef38857ad477f832efdf8816418642024-04-11T18:03:19ZengSAGE PublishingCell Transplantation1555-38922024-04-013310.1177/09636897241242624Optimization of Xenografting Methods for Generating Human Skeletal Muscle in MiceAndrea O’Neill0Anna Llach Martinez1Amber L. Mueller2Weiliang Huang3Anthony Accorsi4Maureen A. Kane5David Eyerman6Robert J. Bloch7Department of Physiology, University of Maryland School of Medicine, Baltimore, MD, USADepartment of Physiology, University of Maryland School of Medicine, Baltimore, MD, USACell Metabolism, Cambridge, MA, USADepartment of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD, USABlackbird Laboratories, Baltimore, MD, USADepartment of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD, USAApellis Pharmaceuticals, Waltham, MA, USADepartment of Physiology, University of Maryland School of Medicine, Baltimore, MD, USAXenografts 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.https://doi.org/10.1177/09636897241242624 |
| spellingShingle | Andrea O’Neill Anna Llach Martinez Amber L. Mueller Weiliang Huang Anthony Accorsi Maureen A. Kane David Eyerman Robert J. Bloch Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice Cell Transplantation |
| title | Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice |
| title_full | Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice |
| title_fullStr | Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice |
| title_full_unstemmed | Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice |
| title_short | Optimization of Xenografting Methods for Generating Human Skeletal Muscle in Mice |
| title_sort | optimization of xenografting methods for generating human skeletal muscle in mice |
| url | https://doi.org/10.1177/09636897241242624 |
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