論文

査読有り 国際誌
2019年10月

Engineering of human cardiac muscle electromechanically matured to an adult-like phenotype.

Nature protocols
  • Kacey Ronaldson-Bouchard
  • Keith Yeager
  • Diogo Teles
  • Timothy Chen
  • Stephen Ma
  • LouJin Song
  • Kumi Morikawa
  • Holly M Wobma
  • Alessandro Vasciaveo
  • Edward C Ruiz
  • Masayuki Yazawa
  • Gordana Vunjak-Novakovic
  • 全て表示

14
10
開始ページ
2781
終了ページ
2817
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1038/s41596-019-0189-8

The application of tissue-engineering approaches to human induced pluripotent stem (hiPS) cells enables the development of physiologically relevant human tissue models for in vitro studies of development, regeneration, and disease. However, the immature phenotype of hiPS-derived cardiomyocytes (hiPS-CMs) limits their utility. We have developed a protocol to generate engineered cardiac tissues from hiPS cells and electromechanically mature them toward an adult-like phenotype. This protocol also provides optimized methods for analyzing these tissues' functionality, ultrastructure, and cellular properties. The approach relies on biological adaptation of cultured tissues subjected to biomimetic cues, applied at an increasing intensity, to drive accelerated maturation. hiPS cells are differentiated into cardiomyocytes and used immediately after the first contractions are observed, when they still have developmental plasticity. This starting cell population is combined with human dermal fibroblasts, encapsulated in a fibrin hydrogel and allowed to compact under passive tension in a custom-designed bioreactor. After 7 d of tissue formation, the engineered tissues are matured for an additional 21 d by increasingly intense electromechanical stimulation. Tissue properties can be evaluated by measuring contractile function, responsiveness to electrical stimuli, ultrastructure properties (sarcomere length, mitochondrial density, networks of transverse tubules), force-frequency and force-length relationships, calcium handling, and responses to β-adrenergic agonists. Cell properties can be evaluated by monitoring gene/protein expression, oxidative metabolism, and electrophysiology. The protocol takes 4 weeks and requires experience in advanced cell culture and machining methods for bioreactor fabrication. We anticipate that this protocol will improve modeling of cardiac diseases and testing of drugs.

リンク情報
DOI
https://doi.org/10.1038/s41596-019-0189-8
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/31492957
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7195192
ID情報
  • DOI : 10.1038/s41596-019-0189-8
  • PubMed ID : 31492957
  • PubMed Central 記事ID : PMC7195192

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