Abstract
Cardiomyocytes (CMs) derived from induced pluripotent stem cells (iPSCs) hold great promise for patient-specific disease modeling, drug screening and cell therapy. However, existing protocols for CM differentiation of iPSCs besides being highly dependent on the application of expensive growth factors show low reproducibility and scalability. The aim of this work was to develop a robust and scalable strategy for mass production of iPSC-derived CMs by designing a bioreactor protocol that ensures a hypoxic and mechanical environment. Murine iPSCs were cultivated as aggregates in either stirred tank or WAVE bioreactors. The effect of dissolved oxygen and mechanical forces, promoted by different hydrodynamic environments, on CM differentiation was evaluated. Combining a hypoxia culture (4 % O2 tension) with an intermittent agitation profile in stirred tank bioreactors resulted in an improvement of about 1000-fold in CM yields when compared to normoxic (20 % O2 tension) and continuously agitated cultures. Additionally, we showed for the first time that wave-induced agitation enables the differentiation of iPSCs towards CMs at faster kinetics and with higher yields (60 CMs/input iPSC). In an 11-day differentiation protocol, clinically relevant numbers of CMs (2.3 × 109 CMs/1 L) were produced, and CMs exhibited typical cardiac sarcomeric structures, calcium transients, electrophysiological profiles and drug responsiveness. This work describes significant advances towards scalable cardiomyocyte differentiation of murine iPSC, paving the way for the implementation of this strategy for mass production of their human counterparts and their use for cardiac repair and cardiovascular research.Electronic supplementary materialThe online version of this article (doi:10.1007/s12015-014-9533-0) contains supplementary material, which is available to authorized users.
Highlights
The inability of mature cardiomyocytes (CMs) to proliferate leads to a permanent loss of functional cells after injury [1]
In order to establish a robust and scalable platform for production of functional induced pluripotent stem cells (iPSCs)-derived CMs, we first investigated the effect of dissolved oxygen (DO) on cardiac iPSC differentiation in stirred tank bioreactor systems
In this study we developed a new method for mass production of murine iPSC-derived CMs using environmentally controlled bioreactors
Summary
The inability of mature cardiomyocytes (CMs) to proliferate leads to a permanent loss of functional cells after injury [1]. Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have emerged as an attractive candidate stem cell source for obtaining CMs [3, 4]. The inherent capacity to grow indefinitely and to differentiate into all mature cells of the human body make PSCs the only cell source that can provide ex-vivo an unlimited number of functional and potentially autologous CMs for transplantation. IPSCs can circumvent these drawbacks, allowing for ethically “acceptable” and safe patient-specific therapies [6, 7]. IPSCs constitute a promising tool to establish disease-specific models of human inherited cardiac disorders and platforms for drug discovery and toxicity testing [7, 8]
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