title: Development of a scalable suspension culture for cardiac differentiation from human pluripotent stem cells.
abstract: To meet the need of a large quantity of hPSC-derived cardiomyocytes (CM) for pre-clinical and clinical studies, a robust and scalable differentiation system for CM production is essential. With a human pluripotent stem cells (hPSC) aggregate suspension culture system we established previously, we developed a matrix-free, scalable, and GMP-compliant process for directing hPSC differentiation to CM in suspension culture by modulating Wnt pathways with small molecules. By optimizing critical process parameters including: cell aggregate size, small molecule concentrations, induction timing, and agitation rate, we were able to consistently differentiate hPSCs to >90% CM purity with an average yield of 1.5 to 2×10(9) CM/L at scales up to 1L spinner flasks. CM generated from the suspension culture displayed typical genetic, morphological, and electrophysiological cardiac cell characteristics. This suspension culture system allows seamless transition from hPSC expansion to CM differentiation in a continuous suspension culture. It not only provides a cost and labor effective scalable process for large scale CM production, but also provides a bioreactor prototype for automation of cell manufacturing, which will accelerate the advance of hPSC research towards therapeutic applications.
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Vincent C
authors: VC
authors: Chen
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Jingjing
authors: J
authors: Ye
authors: Center for Applied Technology Development, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
authors: Praveen
authors: P
authors: Shukla
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Giau
authors: G
authors: Hua
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Danlin
authors: D
authors: Chen
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Ziguang
authors: Z
authors: Lin
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Jian-chang
authors: JC
authors: Liu
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Jing
authors: J
authors: Chai
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
authors: Joseph
authors: J
authors: Gold
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
authors: Joseph
authors: J
authors: Wu
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: David
authors: D
authors: Hsu
authors: Center for Biomedicine and Genetics, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA; Center for Applied Technology Development, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, USA.
authors: Larry A
authors: LA
authors: Couture
mesh_terms: D000185:Actinin / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D005786:Gene Expression Regulation; D006801:Humans; D008856:Microscopy, Fluorescence; D032383:Myocytes, Cardiac / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D017423:Sequence Analysis, RNA; D019210:Troponin I / Q000378:metabolism; D020107:Troponin T / Q000378:metabolism; D060449:Wnt Signaling Pathway
chemical_list: D019210:Troponin I; D020107:Troponin T; D000185:Actinin; D012313:RNA
keywords: Cardiomyocyte differentiation; GMP; Human pluripotent stem cells; Suspension cell cultures