SOUIDI, M., Y. SLEIMAN, Ivana AĆIMOVIĆ, Jan PŘIBYL, V. CHARRABI, V. BEACKER, V. SCHEUERMANN, Martin PEŠL, Šárka JELÍNKOVÁ, Petr SKLÁDAL, Petr DVOŘÁK, A. LACAMPAGNE, Vladimír ROTREKL a Albano MELI. Role of oxygen exposure on the differentiation of human induced pluripotent stem cells in 2D and 3D cardiac organoids. 2021. Dostupné z: https://dx.doi.org/10.1016/j.acvdsp.2021.04.204.
Další formáty:   BibTeX LaTeX RIS
Základní údaje
Originální název Role of oxygen exposure on the differentiation of human induced pluripotent stem cells in 2D and 3D cardiac organoids
Autoři SOUIDI, M., Y. SLEIMAN, Ivana AĆIMOVIĆ, Jan PŘIBYL, V. CHARRABI, V. BEACKER, V. SCHEUERMANN, Martin PEŠL, Šárka JELÍNKOVÁ, Petr SKLÁDAL, Petr DVOŘÁK, A. LACAMPAGNE, Vladimír ROTREKL a Albano MELI.
Vydání 2021.
Další údaje
Originální jazyk čeština
Typ výsledku Konferenční abstrakta
Stát vydavatele Česká republika
Utajení není předmětem státního či obchodního tajemství
WWW URL
Organizace Lékařská fakulta – Masarykova univerzita – Repozitář
Doi http://dx.doi.org/10.1016/j.acvdsp.2021.04.204
Změnil Změnil: RNDr. Daniel Jakubík, učo 139797. Změněno: 3. 5. 2022 04:27.
Anotace
Introduction Human induced pluripotent stem cells (hiPSC) have the ability to differentiate theoritically into any cell type. The development of organoid systems exhibiting the essential features of human organ such as liver and heart is of high interest. Optimizing the culture conditions to obtain the highest cardiac organoids efficacy is crucial. In fact, cardiac differentiation protocols have been established by essentially focusing on specific growth factors on hiPSC differentiation efficiency. However, the optimal environmental factors such as the optimal oxygen exposure to obtain cardiac myocytes in network are still unclear. The mesoderm germ layer differentiation is known to be enhanced by low oxygen exposure. Yet, the effect of low oxygen exposure on the molecular and functional maturity of the hiPSC-derived cardiomyocytes remains unexplored. Aims We aimed here at comparing the molecular and functional consequences of low (5% O2 or LOE) and high oxygen exposure (21% O2 or HOE) on cardiac differentiation of hiPSCs in 2D monolayer and 3D organoids protocols. Methods hiPSC-CMs were differentiated through both the 2D (monolayer) and 3D (embryoid body) protocols using several lines. Cardiac marker expression and cell morphology were assessed using qRT-PCR and immunofluorescence. The mitochondrial localization and metabolic properties were evaluated by high-resolution respirometry and mitochondrial staining. The intracellular Ca2+ handling and contractile properties were also monitored using confocal fluorescent microscopy and atomic force microscopy. Results Our results indicated that the 2D cardiac monolayer can only be differentiated in HOE. The 3D cardiac organoids containing hiPSC-CMs in LOE exhibited higher cardiac markers expression such as troponin T (TnTc), RyR2, Serca2a, alpha and beta heavy myosin chains. Moreover, we found enhanced contractile force, hypertrophy and steadier SR Ca2+ release reflected by a more regular spontaneous Ca2+ transients associated with a higher maximal amplitude and lower spontaneous Ca2+ events revealing a better SR Ca2+ handling in LOE. Similar beat rate, preserved distribution of mitochondria and similar oxygen consumption by the mitochondrial respiratory chain complexes were also observed. Conclusions Our results brought evidences that LOE is moderately beneficial for the 3D cardiac organoids with hPSC-CMs exhibiting further maturity. In contrast, the 2D cardiac monolayers strictly require HOE.
Anotace anglicky
Introduction Human induced pluripotent stem cells (hiPSC) have the ability to differentiate theoritically into any cell type. The development of organoid systems exhibiting the essential features of human organ such as liver and heart is of high interest. Optimizing the culture conditions to obtain the highest cardiac organoids efficacy is crucial. In fact, cardiac differentiation protocols have been established by essentially focusing on specific growth factors on hiPSC differentiation efficiency. However, the optimal environmental factors such as the optimal oxygen exposure to obtain cardiac myocytes in network are still unclear. The mesoderm germ layer differentiation is known to be enhanced by low oxygen exposure. Yet, the effect of low oxygen exposure on the molecular and functional maturity of the hiPSC-derived cardiomyocytes remains unexplored. Aims We aimed here at comparing the molecular and functional consequences of low (5% O2 or LOE) and high oxygen exposure (21% O2 or HOE) on cardiac differentiation of hiPSCs in 2D monolayer and 3D organoids protocols. Methods hiPSC-CMs were differentiated through both the 2D (monolayer) and 3D (embryoid body) protocols using several lines. Cardiac marker expression and cell morphology were assessed using qRT-PCR and immunofluorescence. The mitochondrial localization and metabolic properties were evaluated by high-resolution respirometry and mitochondrial staining. The intracellular Ca2+ handling and contractile properties were also monitored using confocal fluorescent microscopy and atomic force microscopy. Results Our results indicated that the 2D cardiac monolayer can only be differentiated in HOE. The 3D cardiac organoids containing hiPSC-CMs in LOE exhibited higher cardiac markers expression such as troponin T (TnTc), RyR2, Serca2a, alpha and beta heavy myosin chains. Moreover, we found enhanced contractile force, hypertrophy and steadier SR Ca2+ release reflected by a more regular spontaneous Ca2+ transients associated with a higher maximal amplitude and lower spontaneous Ca2+ events revealing a better SR Ca2+ handling in LOE. Similar beat rate, preserved distribution of mitochondria and similar oxygen consumption by the mitochondrial respiratory chain complexes were also observed. Conclusions Our results brought evidences that LOE is moderately beneficial for the 3D cardiac organoids with hPSC-CMs exhibiting further maturity. In contrast, the 2D cardiac monolayers strictly require HOE.
Typ Název Vložil/a Vloženo Práva
1-s2.0-S1878648021002068-main.pdf Licence Creative Commons  Verze souboru 25. 1. 2022

Vlastnosti

Název
1-s2.0-S1878648021002068-main.pdf
Adresa v ISu
https://repozitar.cz/auth/repo/48274/1232802/
Adresa ze světa
https://repozitar.cz/repo/48274/1232802/
Adresa do Správce
https://repozitar.cz/auth/repo/48274/1232802/?info
Ze světa do Správce
https://repozitar.cz/repo/48274/1232802/?info
Vloženo
Út 25. 1. 2022 14:41

Práva

Právo číst
  • kdokoliv v Internetu
Právo vkládat
 
Právo spravovat
  • osoba Mgr. Lucie Vařechová, uco 106253
  • osoba RNDr. Daniel Jakubík, uco 139797
  • osoba Mgr. Jolana Surýnková, uco 220973
Atributy
 
Vytisknout
Přidat do schránky Zobrazeno: 17. 5. 2024 11:53