Practical adoption of state-of-the-art hiPSC-cardiomyocyte differentiation techniques.

PloS One
Cassady E RupertKareen L K Coulombe

Abstract

Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are a valuable resource for cardiac therapeutic development; however, generation of these cells in large numbers and high purity is a limitation in widespread adoption. Here, design of experiments (DOE) is used to investigate the cardiac differentiation space of three hiPSC lines when varying CHIR99027 concentration and cell seeding density, and a novel image analysis is developed to evaluate plate coverage when initiating differentiation. Metabolic selection via lactate purifies hiPSC-cardiomyocyte populations, and the bioenergetic phenotype and engineered tissue mechanics of purified and unpurified hiPSC-cardiomyocytes are compared. Findings demonstrate that when initiating differentiation one day after hiPSC plating, low (3 μM) Chiron and 72 x 103 cells/cm2 seeding density result in peak cardiac purity (50-90%) for all three hiPSC lines. Our results confirm that metabolic selection with lactate shifts hiPSC-cardiomyocyte metabolism towards oxidative phosphorylation, but this more "mature" metabolic phenotype does not by itself result in a more mature contractile phenotype in engineered cardiac tissues at one week of culture in 3D tissues. This study provides ...Continue Reading

References

Apr 25, 2000·Journal of Molecular and Cellular Cardiology·P A DoevendansR S Kass
Jul 11, 2008·Pediatric Research·Michael A YatscoffGary D Lopaschuk
Nov 4, 2009·Trends in Biochemical Sciences·Dianqing Wu, Weijun Pan
Feb 15, 2011·The Journal of Clinical Investigation·Kazim H NarsinhJoseph C Wu
Mar 9, 2012·Regenerative Medicine·Brian LiauNenad Bursac
May 31, 2012·Proceedings of the National Academy of Sciences of the United States of America·Xiaojun LianSean P Palecek
Jul 31, 2012·Toxicological Sciences : an Official Journal of the Society of Toxicology·Payal RanaYvonne Will
Jun 16, 2014·Nature Methods·Paul W BurridgeJoseph C Wu
Jul 1, 2015·Nature Methods·Xiaojun LianSean P Palecek
Oct 7, 2015·Current Protocols in Human Genetics·Paul W BurridgeJoseph C Wu
Jan 31, 2016·Cell Adhesion & Migration·Martin Gauster, Berthold Huppertz
May 2, 2017·Tissue Engineering. Part C, Methods·Fabiola MunarinKareen L K Coulombe
Jul 8, 2017·Circulation·Thomas EschenhagenJoseph A Hill
Nov 28, 2018·Communications Biology·Ngan F HuangJoseph C Wu

❮ Previous
Next ❯

Citations

Sep 6, 2020·Animals : an Open Access Journal From MDPI·Alexandra Bizy, Matthew Klos
Jun 16, 2021·Journal of the Mechanical Behavior of Biomedical Materials·Yifei BaiVikas Srivastava

❮ Previous
Next ❯

Datasets Mentioned

BETA
GM25256

Methods Mentioned

BETA
flow cytometry

Software Mentioned

MathWorks
Graphpad Prism
Flowing
Chiron
MATLAB
DOE
IonOptix

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.