Hamad, Sarkawt ORCID: 0000-0002-5155-9467, Derichsweiler, Daniel, Papadopoulos, Symeon, Nguemo, Filomain, Saric, Tomo, Sachinidis, Agapios, Brockmeier, Konrad, Hescheler, Juergen, Boukens, Bastiaan J. and Pfannkuche, Kurt (2019). Generation of human induced pluripotent stem cell-derived cardiomyocytes in 2D monolayer and scalable 3D suspension bioreactor cultures with reduced batch-to-batch variations. Theranostics, 9 (24). S. 7222 - 7239. LAKE HAVEN: IVYSPRING INT PUBL. ISSN 1838-7640

Full text not available from this repository.

Abstract

Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) are promising candidates to treat myocardial infarction and other cardiac diseases. Such treatments require pure cardiomyocytes (CMs) in large quantities. Methods: In the present study we describe an improved protocol for production of hiPSC-CMs in which hiPSCs are first converted into mesodermal cells by stimulation of wingless (Wnt) signaling using CHIR99021, which are then further differentiated into CM progenitors by simultaneous inhibition of porcupine and tankyrase pathways using IWP2 and XAV939 under continuous supplementation of ascorbate during the entire differentiation procedure. Results: The protocol resulted in reproducible generation of >90% cardiac troponin T (TNNT2)-positive cells containing highly organized sarcomeres. In 2D monolayer cultures CM yields amounted to 0.5 million cells per cm 2 growth area, and on average 72 million cells per 100 mL bioreactor suspension culture without continuous perfusion. The differentiation efficiency was hardly affected by the initial seeding density of undifferentiated hiPSCs. Furthermore, batch-to-batch variations were reduced by combinatorial use of ascorbate, IWP2, and XAV939. Conclusion: Combined inhibition of porcupine and tankyrase sub-pathways of Wnt signaling and continuous ascorbate supplementation, enable robust and efficient production of hiPSC-CMs.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Hamad, SarkawtUNSPECIFIEDorcid.org/0000-0002-5155-9467UNSPECIFIED
Derichsweiler, DanielUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Papadopoulos, SymeonUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Nguemo, FilomainUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Saric, TomoUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Sachinidis, AgapiosUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Brockmeier, KonradUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hescheler, JuergenUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Boukens, Bastiaan J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Pfannkuche, KurtUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-160558
DOI: 10.7150/thno.32058
Journal or Publication Title: Theranostics
Volume: 9
Number: 24
Page Range: S. 7222 - 7239
Date: 2019
Publisher: IVYSPRING INT PUBL
Place of Publication: LAKE HAVEN
ISSN: 1838-7640
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
CARDIAC DIFFERENTIATION; WNT; ANTAGONISMMultiple languages
Medicine, Research & ExperimentalMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/16055

Downloads

Downloads per month over past year

Altmetric

Export

Actions (login required)

View Item View Item