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.

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).

Reviewing Editor

  1. Amy J Wagers, Harvard University, United States

Publication 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

  • 18,814
    Page views
  • 2,030
    Downloads
  • 192
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, 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
  1. Further reading

Further reading

    1. Stem Cells and Regenerative Medicine
    Jianhua Zhang et al.
    Research Article

    Research and therapeutic applications using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) require robust differentiation strategies. Efforts to improve hPSC-CM differentiation have largely overlooked the role of extracellular matrix (ECM). The present study investigates the ability of defined ECM proteins to promote hPSC cardiac differentiation. Fibronectin (FN), laminin-111, and laminin-521 enabled hPSCs to attach and expand. However, only addition of FN promoted cardiac differentiation in response to growth factors Activin A, BMP4, and bFGF in contrast to the inhibition produced by laminin-111 or laminin-521. hPSCs in culture produced endogenous FN which accumulated in the ECM to a critical level necessary for effective cardiac differentiation. Inducible shRNA knockdown of FN prevented Brachyury+ mesoderm formation and subsequent hPSC-CM generation. Antibodies blocking FN binding integrins α4β1 or αVβ1, but not α5β1, inhibited cardiac differentiation. Furthermore, inhibition of integrin-linked kinase led to a decrease in phosphorylated AKT, which was associated with increased apoptosis and inhibition of cardiac differentiation. These results provide new insights into defined matrices for culture of hPSCs that enable production of FN-enriched ECM which is essential for mesoderm formation and efficient cardiac differentiation.

    1. Neuroscience
    2. Stem Cells and Regenerative Medicine
    Weizhao Chen et al.
    Short Report Updated

    Lineage reprogramming of resident glial cells to dopaminergic neurons (DAns) is an attractive prospect of the cell-replacement therapy for Parkinson’s disease (PD). However, it is unclear whether repressing polypyrimidine tract binding protein 1 (PTBP1) could efficiently convert astrocyte to DAns in the substantia nigra and striatum. Although reporter-positive DAns were observed in both groups after delivering the adeno-associated virus (AAV) expressing a reporter with shRNA or CRISPR-CasRx to repress astroglial PTBP1, the possibility of AAV leaking into endogenous DAns could not be excluded without using a reliable lineage-tracing method. By adopting stringent lineage-tracing strategy, two other studies show that either knockdown or genetic deletion of quiescent astroglial PTBP1 fails to obtain induced DAns under physiological condition. However, the role of reactive astrocytes might be underestimated because upon brain injury, reactive astrocyte can acquire certain stem cell hallmarks that may facilitate the lineage conversion process. Therefore, whether reactive astrocytes could be genuinely converted to DAns after PTBP1 repression in a PD model needs further validation. In this study, we used Aldh1l1-CreERT2-mediated specific astrocyte-lineage-tracing method to investigate whether reactive astrocytes could be converted to DAns in a 6-hydroxydopamine (6-OHDA) mouse model of PD. However, we found that no astrocyte-originated DAn was generated after effective and persistent knockdown of astroglial PTBP1 either in the substantia nigra or in striatum, while AAV ‘leakage’ to nearby neurons was easily observed. Our results confirm that repressing PTBP1 does not convert astrocytes to DAns, regardless of physiological or PD-related pathological conditions.