A complete workflow for the differentiation and the dissociation of hiPSC-derived cardiospheres - ScienceDirect

Stem Cell Research

Benjamin Fischer^1¹  Anna Meier^1,2²  Sascha Neumann^1,4³  Annika Dehne^1⁴  Aseem Salhotra^1,3⁵  Thao Anh Tran^1⁶ 

[1] Fraunhofer Institute for Biomedical Engineering, Joseph-von-Fraunhofer-Weg 1, Sulzbach 66280, Germany^1;Fraunhofer Project Centre for Stem Cell Process Engineering, Neunerplatz 2, Würzburg 97082, Germany^2;Hasselt University, Campus Diepenbeek, Diepenbeek 3590, Belgium^6;Saarland University, Gebäude A, Saarbrücken 66123, Germany^3;Universidad Católica del Norte, Larrondo 1281, Coquimbo 1780000, Chile^4;University of Applied Sciences Kaiserslautern, Campus Zweibrücken, Building G, Amerikastr. 1, Zweibrücken 66482, Germany^5

关键词: hiPSC;    human induced pluripotent stem cell;    CM;    cardiomyocyte;    BL;    BioLevitator;    HD;    hanging drop;    NC;    NucleoCounter;    LT;    LeviTube;    Ca;    Calcein;    hiPSCs;    Cardiomyocytes;    Cardiac induction;    3D bioreactor;    Papain dissociation;   

DOI  :  10.1016/j.scr.2018.08.015

学科分类：生物科学（综合）

来源: Elsevier

【 摘 要 】

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) are an invaluable tool for both basic and translational cardiovascular research. The potential that these cells hold for therapy, disease modeling and drug discovery is hampered by several bottlenecks that currently limit both the yield and the efficiency of cardiac induction. Here, we present a complete workflow for the production of ready-to-use hiPSC-CMs in a dynamic suspension bioreactor. This includes the efficient and highly reproducible differentiation of hiPSCs into cardiospheres, which display enhanced physiological maturation compared to static 3D induction in hanging drops, and a novel papain-based dissociation method that offers higher yield and viability than the broadly used dissociation reagents TrypLE and Accutase. Molecular and functional analyses of the cardiomyocytes reseeded after dissociation confirmed both the identity and the functionality of the cells, which can be used in downstream applications, either as monolayers or spheroids.

【 授权许可】

Unknown