title: Universal cardiac induction of human pluripotent stem cells in two and three-dimensional formats: implications for in vitro maturation.
abstract: Directed cardiac differentiation of human pluripotent stem cells (hPSCs) enables disease modeling, investigation of human cardiogenesis, as well as large-scale production of cardiomyocytes (CMs) for translational purposes. Multiple CM differentiation protocols have been developed to individually address specific requirements of these diverse applications, such as enhanced purity at a small scale or mass production at a larger scale. However, there is no universal high-efficiency procedure for generating CMs both in two-dimensional (2D) and three-dimensional (3D) culture formats, and undefined or complex media additives compromise functional analysis or cost-efficient upscaling. Using systematic combinatorial optimization, we have narrowed down the key requirements for efficient cardiac induction of hPSCs. This implied differentiation in simple serum and serum albumin-free basal media, mediated by a minimal set of signaling pathway manipulations at moderate factor concentrations. The method was applicable both to 2D and 3D culture formats as well as to independent hPSC lines. Global time-course gene expression analyses over extended time periods and in comparison with human heart tissue were used to monitor culture-induced maturation of the resulting CMs. This suggested that hPSC-CMs obtained with our procedure reach a rather stable transcriptomic state after approximately 4 weeks of culture. The underlying gene expression changes correlated well with a decline of immature characteristics as well as with a gain of structural and physiological maturation features within this time frame. These data link gene expression patterns of hPSC-CMs to functional readouts and thus define the cornerstones of culture-induced maturation.
authors: Human Stem Cell Pluripotency Group; Chemical Genomics Centre of the Max Planck Society, Dortmund, Germany.
authors: Miao
authors: M
authors: Zhang
authors: Jan Sebastian
authors: JS
authors: Schulte
authors: Alexander
authors: A
authors: Heinick
authors: Ilaria
authors: I
authors: Piccini
authors: Jyoti
authors: J
authors: Rao
authors: Roberto
authors: R
authors: Quaranta
authors: Dagmar
authors: D
authors: Zeuschner
authors: Daniela
authors: D
authors: Malan
authors: Kee-Pyo
authors: KP
authors: Kim
authors: Albrecht
authors: A
authors: Röpke
authors: Philipp
authors: P
authors: Sasse
authors: Marcos
authors: M
authors: Araúzo-Bravo
authors: Guiscard
authors: G
authors: Seebohm
authors: Hans
authors: H
authors: Schöler
authors: Larissa
authors: L
authors: Fabritz
authors: Paulus
authors: P
authors: Kirchhof
authors: Frank Ulrich
authors: FU
authors: Müller
authors: Boris
authors: B
authors: Greber
mesh_terms: D002454:Cell Differentiation; D006321:Heart / Q000502:physiology*; D006801:Humans; D008648:Mesoderm / Q000166:cytology; D032383:Myocytes, Cardiac / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology*
keywords: cardiac differentiation; human pluripotent stem cells; in-vitro maturation