J 2014

Forced aggregation and defined factors allow highly uniform-sized embryoid bodies and functional cardiomyocytes from human embryonic and induced pluripotent stem cells

PEŠL, Martin, Ivana AĆIMOVIĆ, Jan PŘIBYL, Renata HÉŽOVÁ, Aleksandra VILOTIĆ et. al.

Basic information

Original name

Forced aggregation and defined factors allow highly uniform-sized embryoid bodies and functional cardiomyocytes from human embryonic and induced pluripotent stem cells

Authors

PEŠL, Martin (203 Czech Republic, guarantor, belonging to the institution), Ivana AĆIMOVIĆ (688 Serbia, belonging to the institution), Jan PŘIBYL (203 Czech Republic, belonging to the institution), Renata HÉŽOVÁ (203 Czech Republic, belonging to the institution), Aleksandra VILOTIĆ (688 Serbia, belonging to the institution), Jeremy FAUCONNIER (250 France), Jan VRBSKÝ (203 Czech Republic, belonging to the institution), Peter KRUZLIAK (203 Czech Republic), Petr SKLÁDAL (203 Czech Republic, belonging to the institution), Tomáš KÁRA (203 Czech Republic), Vladimír ROTREKL (203 Czech Republic, belonging to the institution), Alain LACAMPAGNE (250 France), Petr DVOŘÁK (203 Czech Republic, belonging to the institution) and Albano MELI (250 France, belonging to the institution)

Edition

Heart Vessels, Japan, Springer Japan, 2014, 0910-8327

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

Japan

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 2.065

RIV identification code

RIV/00216224:14110/14:00074888

Organization unit

Faculty of Medicine

UT WoS

000345145200014

Keywords (in Czech)

human pluripotent stem cell embryoid body differentiation cardiomyocyte calcium

Keywords in English

human pluripotent stem cell embryoid body differentiation cardiomyocyte calcium

Tags

Tags

International impact, Reviewed
Změněno: 15/1/2015 12:06, Ing. Mgr. Věra Pospíšilíková

Abstract

V originále

In vitro human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) can differentiate into functional cardiomyocytes (CMs). Protocols for cardiac differentiation of hESCs and hiPSCs include formation of the three-dimensional cell aggregates called embryoid bodies (EBs). The traditional suspension method for EB formation from clumps of cells results in an EB population heterogeneous in size and shape. In this study we show that forced aggregation of a defined number of single cells on AggreWell plates gives a high number of homogeneous EBs that can be efficiently differentiated into functional CMs by application of defined growth factors in the media. For cardiac differentiation, we used three hESC lines and one hiPSC line. Our contracting EBs and the resulting CMs express cardiac markers, namely myosin heavy chain alpha and beta, cardiac ryanodine receptor/calcium release channel, and cardiac troponin T, shown by real-time polymerase chain reaction and immunocytochemistry. Using Ca2+ imaging and atomic force microscopy, we demonstrate the functionality of RyR2 to release Ca2+ from the sarcoplasmic reticulum as well as reliability in contractile and beating properties of hESC-EBs and hiPSC-EBs upon the stimulation or inhibition of the beta-adrenergic pathway.

Links

ED1.1.00/02.0068, research and development project
Name: CEITEC - central european institute of technology
EE2.3.20.0011, research and development project
Name: Centrum výzkumu pluripotentních buněk a nestability genomu
MSM0021622430, plan (intention)
Name: Funkční a molekulární charakteristiky nádorových a normálních kmenových buněk - identifikace cílů pro nová terapeutika a terapeutické strategie
Investor: Ministry of Education, Youth and Sports of the CR, Functional and molecular characteristics of cancer and normal stem cells - identification of targets for novel therapeutics and therapeutic strategies
7AMB13FR011, research and development project
Name: Přeprogramování somatických buněk darovaných pacienty s dědičnou Duchennovou svalovou dystrofií do kardiomyocytů - nahlédnutí do molekulární podstaty patologických dějů u dilatační kardiomyopatie nemocných DMD (Acronym: DUCHENSTEM)
Investor: Ministry of Education, Youth and Sports of the CR

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