title: The effect of microgrooved culture substrates on calcium cycling of cardiac myocytes derived from human induced pluripotent stem cells.
abstract: Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) have been widely proposed as in vitro models of myocardial physiology and disease. A significant obstacle, however, is their immature phenotype. We hypothesised that Ca(2+) cycling of iPSC-CM is influenced by culture conditions and can be manipulated to obtain a more mature cellular behaviour. To test this hypothesis we seeded iPSC-CM onto fibronectin coated microgrooved polydimethylsiloxane (PDMS) scaffolds fabricated using photolithography, or onto unstructured PDMS membrane. After two weeks in culture, the structure and function of iPSC-CM were studied. PDMS microgrooved culture substrates brought about cellular alignment (p < 0.0001) and more organised sarcomere. The Ca(2+) cycling properties of iPSC-CM cultured on these substrates were significantly altered with a shorter time to peak amplitude (p = 0.0002 at 1 Hz), and more organised sarcoplasmic reticulum (SR) Ca(2+) release in response to caffeine (p < 0.0001), suggesting improved SR Ca(2+) cycling. These changes were not associated with modifications in gene expression. Whilst structured tissue culture may make iPSC-CM more representative of adult myocardium, further construct development and characterisation is required to optimise iPSC-CM as a model of adult myocardium.
authors.affiliation: National Heart and Lung Institute, Imperial College London, London, United Kingdom.
authors.forename: Christopher
authors.initials: C
authors.lastname: Rao
authors.forename: Themistoklis
authors.initials: T
authors.lastname: Prodromakis
authors.forename: Ljudmila
authors.initials: L
authors.lastname: Kolker
authors.forename: Umar A R
authors.initials: UA
authors.lastname: Chaudhry
authors.forename: Tatiana
authors.initials: T
authors.lastname: Trantidou
authors.forename: Arun
authors.initials: A
authors.lastname: Sridhar
authors.forename: Claire
authors.initials: C
authors.lastname: Weekes
authors.forename: Patrizia
authors.initials: P
authors.lastname: Camelliti
authors.forename: Sian E
authors.initials: SE
authors.lastname: Harding
authors.forename: Ara
authors.initials: A
authors.lastname: Darzi
authors.forename: Magdi H
authors.initials: MH
authors.lastname: Yacoub
authors.forename: Thanos
authors.initials: T
authors.lastname: Athanasiou
authors.forename: Cesare M
authors.initials: CM
authors.lastname: Terracciano
mesh_terms: D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004594:Electrophysiology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008962:Models, Theoretical; D032383:Myocytes, Cardiac / Q000166:cytology*
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon