Abstract
Human induced pluripotent stem cells (hiPSCs) represent an almost limitless source of cells for disease modelling and drug screening applications. Here we established an efficient and robust 3D platform for cardiomyocyte (CMs) production from hiPSCs, solely through small-molecule-based temporal modulation of the Wnt signalling, which generates more than 90% cTNT+ cells. The impact of performing the differentiation process in 3D conditions as compared to a 2D culture system, was characterized by transcriptomic analysis by using data collected from sequential stages of 2D and 3D culture. We highlight that performing an initial period of hiPSC aggregation before cardiac differentiation primed hiPSCs towards an earlier mesendoderm lineage differentiation, via TGF-β/Nodal signaling stabilization. Importantly, it was also found that CMs in the 3D microenvironment mature earlier and show an improved communication system, which we suggested to be responsible for a higher structural and functional maturation of 3D cardiac aggregates.
Highlights
Aiming at better mimicking the microenvironment of in vivo heart development, 3D platforms for in vitro cardiac differentiation and maturation have emerged in the past few years
To develop a platform for 3D cardiac differentiation of Human induced pluripotent stem cells (hiPSCs), we used the temporal modulation of Wnt signalling pathway[18] and a factorial design approach[19] for the optimization process
The cardiac differentiation process was optimized by testing the combined effect of small molecule CHIR concentration, ranging from 4 μM to 18 μM, and aggregate diameter at day 0, ranging from 220 μm to 360 μm, with the model centred in the culture condition corresponding to 11 μM CHIR and 300 μm, respectively
Summary
We established an efficient and robust 3D platform for cardiomyocyte (CMs) production from hiPSCs, solely through small-molecule-based temporal modulation of the Wnt signalling, which generates more than 90% cTNT+ cells. We expanded the knowledge regarding the impact of 3D culture of hiPSCs in a forced aggregation platform and took advantage of that knowledge to develop a simple, efficient and robust 3D platform for hiPSC differentiation towards CMs, using the temporal modulation of the Wnt signalling pathway. Expression profiling data analyses revealed that the initial period of hiPSC 3D aggregation before cardiac differentiation induces significant transcriptional changes that favour the cardiac differentiation process by priming hiPSCs to mesendoderm lineages. The obtained data suggests that the CMs obtained in this 3D microenvironment mature earlier when compared with 2D cardiac monolayer
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
