Bioengineered human myobundles mimic clinical responses of skeletal muscle to drugs

  1. Lauran Madden
  2. Mark Juhas
  3. William E Kraus
  4. George A Truskey
  5. Nenad Bursac  Is a corresponding author
  1. Duke University, United States
  2. Duke University School of Medicine, United States

Abstract

Existing in vitro models of human skeletal muscle cannot recapitulate the organization and function of native muscle, limiting their use in physiological and pharmacological studies. Here, we demonstrate engineering of electrically and chemically responsive, contractile human muscle tissues ('myobundles') using primary myogenic cells. These biomimetic constructs exhibit aligned architecture, multinucleated and striated myofibers, and a Pax7+ cell pool. They contract spontaneously and respond to electrical stimuli with twitch and tetanic contractions. Positive correlation between contractile force and GCaMP6-reported calcium responses enables non-invasive tracking of myobundle function and drug response. During culture, myobundles maintain functional acetylcholine receptors and structurally and functionally mature, evidenced by increased myofiber diameter and improved calcium handling and contractile strength. In response to diversely acting drugs, myobundles undergo dose-dependent hypertrophy or toxic myopathy similar to clinical outcomes. Human myobundles provide an enabling platform for predictive drug and toxicology screening and development of novel therapeutics for muscle-related disorders.

Article and author information

Author details

  1. Lauran Madden

    Department of Biomedical Engineering, Duke University, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Mark Juhas

    Department of Biomedical Engineering, Duke University, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. William E Kraus

    Department of Medicine, Duke University School of Medicine, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. George A Truskey

    Department of Biomedical Engineering, Duke University, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Nenad Bursac

    Department of Biomedical Engineering, Duke University, Durham, United States
    For correspondence
    nbursac@duke.edu
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Amy J Wagers, Harvard University, United States

Ethics

Human subjects: Human skeletal muscle samples were obtained through standard needle biopsy or surgical waste under Duke University IRB approved protocols (Pro00048509 and Pro00012628).

Version history

  1. Received: September 23, 2014
  2. Accepted: January 8, 2015
  3. Accepted Manuscript published: January 9, 2015 (version 1)
  4. Version of Record published: February 16, 2015 (version 2)

Copyright

© 2015, Madden et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 19,094
    Page views
  • 2,233
    Downloads
  • 240
    Citations

Article citation count generated by polling the highest count across the following sources: Scopus, Crossref, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Lauran Madden
  2. Mark Juhas
  3. William E Kraus
  4. George A Truskey
  5. Nenad Bursac
(2015)
Bioengineered human myobundles mimic clinical responses of skeletal muscle to drugs
eLife 4:e04885.
https://doi.org/10.7554/eLife.04885

Share this article

https://doi.org/10.7554/eLife.04885

Further reading

    1. Stem Cells and Regenerative Medicine
    Qinyu Zhang, Rasmus Olofzon ... David Bryder
    Research Article

    The scarcity of hematopoietic stem cells (HSCs) restricts their use in both clinical settings and experimental research. Here, we examined a recently developed method for expanding rigorously purified murine HSCs ex vivo. After 3 weeks of culture, only 0.1% of cells exhibited the input HSC phenotype, but these accounted for almost all functional long-term HSC activity. Input HSCs displayed varying potential for ex vivo self-renewal, with alternative outcomes revealed by single-cell multimodal RNA and ATAC sequencing profiling. While most HSC progeny offered only transient in vivo reconstitution, these cells efficiently rescued mice from lethal myeloablation. The amplification of functional HSC activity allowed for long-term multilineage engraftment in unconditioned hosts that associated with a return of HSCs to quiescence. Thereby, our findings identify several key considerations for ex vivo HSC expansion, with major implications also for assessment of normal HSC activity.

    1. Stem Cells and Regenerative Medicine
    Matias I Autio, Efthymios Motakis ... Roger SY Foo
    Research Article Updated

    Selection of the target site is an inherent question for any project aiming for directed transgene integration. Genomic safe harbour (GSH) loci have been proposed as safe sites in the human genome for transgene integration. Although several sites have been characterised for transgene integration in the literature, most of these do not meet criteria set out for a GSH and the limited set that do have not been characterised extensively. Here, we conducted a computational analysis using publicly available data to identify 25 unique putative GSH loci that reside in active chromosomal compartments. We validated stable transgene expression and minimal disruption of the native transcriptome in three GSH sites in vitro using human embryonic stem cells (hESCs) and their differentiated progeny. Furthermore, for easy targeted transgene expression, we have engineered constitutive landing pad expression constructs into the three validated GSH in hESCs.