Large-scale production of cardiomyocytes from human pluripotent stem cells using a highly reproducible small molecule-based differentiation protocol
File version
Version of Record (VoR)
Author(s)
Ansari, H
Abbasalizadeh, S
Blue, GM
Aghdami, N
Winlaw, DS
Harvey, RP
Bosman, A
Baharvand, H
Griffith University Author(s)
Primary Supervisor
Other Supervisors
Editor(s)
Date
Size
File type(s)
Location
License
Abstract
Maximizing the benefit of human pluripotent stem cells (hPSCs) for research, disease modeling, pharmaceutical and clinical applications requires robust methods for the large-scale production of functional cell types, including cardiomyocytes. Here we demonstrate that the temporal manipulation of WNT, TGF-β, and SHH signaling pathways leads to highly efficient cardiomyocyte differentiation of single-cell passaged hPSC lines in both static suspension and stirred suspension bioreactor systems. Employing this strategy resulted in ~ 100% beating spheroids, consistently containing > 80% cardiac troponin T-positive cells after 15 days of culture, validated in multiple hPSC lines. We also report on a variation of this protocol for use with cell lines not currently adapted to single-cell passaging, the success of which has been verified in 42 hPSC lines. Cardiomyocytes generated using these protocols express lineage-specific markers and show expected electrophysiological functionalities. Our protocol presents a simple, efficient and robust platform for the large-scale production of human cardiomyocytes.
Journal Title
Journal of Visualized Experiments
Conference Title
Book Title
Edition
Volume
Issue
113
Thesis Type
Degree Program
School
Publisher link
DOI
Patent number
Funder(s)
Grant identifier(s)
Rights Statement
Rights Statement
© 2016 The Authors. The attached file is reproduced here in accordance with the copyright policy of the publisher. Please refer to the journal's website for access to the definitive, published version.
Item Access Status
Note
Access the data
Related item(s)
Subject
Biochemistry and cell biology
Psychology
Cognitive and computational psychology
Persistent link to this record
Citation
Fonoudi, H; Ansari, H; Abbasalizadeh, S; Blue, GM; Aghdami, N; Winlaw, DS; Harvey, RP; Bosman, A; Baharvand, H, Large-scale production of cardiomyocytes from human pluripotent stem cells using a highly reproducible small molecule-based differentiation protocol, Journal of Visualized Experiments, 2016, (113), pp. e54276