16332
2018
eng
preprint
1
2018-06-22
--
--
Metabolic substrate shift in human induced pluripotent stem cells during cardiac differentiation: Functional assessment using in vitro radionuclide uptake assay
Background: Recent developments in cellular reprogramming technology enable the production of virtually unlimited numbers of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM). Although hiPSC-CM share various characteristic hallmarks with endogenous cardiomyocytes, it remains a question as to what extent metabolic characteristics are equivalent to mature mammalian cardiomyocytes. Here we set out to functionally characterize the metabolic status of hiPSC-CM in vitro by employing a radionuclide tracer uptake assay. Material and Methods: Cardiac differentiation of hiPSC was induced using a combination of well-orchestrated extrinsic stimuli such as WNT activation (by CHIR99021) and BMP signalling followed by WNT inhibition and lactate based cardiomyocyte enrichment. For characterization of metabolic substrates, dual tracer uptake studies were performed with \(^{18}\)F-2-fluoro-2-deoxy-D-glucose (\(^{18}\)F-FDG) and \(^{125}\)I-β-methyl-iodophenyl-pentadecanoic acid (\(^{125}\)I-BMIPP) as transport markers of glucose and fatty acids, respectively. Results: After cardiac differentiation of hiPSC, in vitro tracer uptake assays confirmed metabolic substrate shift from glucose to fatty acids that was comparable to those observed in native isolated human cardiomyocytes. Immunostaining further confirmed expression of fatty acid transport and binding proteins on hiPSC-CM. Conclusions: During in vitro cardiac maturation, we observed a metabolic shift to fatty acids, which are known as a main energy source of mammalian hearts, suggesting hi-PSC-CM as a potential functional phenotype to investigate alteration of cardiac metabolism in cardiac diseases. Results also highlight the use of available clinical nuclear medicine tracers as functional assays in stem cell research for improved generation of autologous differentiated cells for numerous biomedical applications.
International Journal of Cardiology
10.1016/j.ijcard.2018.06.089
0167-5273
urn:nbn:de:bvb:20-opus-163320
Johns Hopkins School of Medicine, The Russell H Morgan Department of Radiology and Radiological Science, Baltimore, MD, USA
Department of Biomedical Imaging, National Cerebral and Cardiovascular Research Center, Suita, Japan
Institut for Molecular Biology and CMBI, Department of Genomics, Stem Cell Biology and Regenerative Medicine, Leopold-Franzens-University Innsbruck, Innsbruck, Austria
Division of Medical Technology and Science, Department of Medical Physics and Engineering, Course of Health Science, Osaka University Graduate School of Medicine, Suita Japan
701983
Naoko Nose, Rudolf A. Werner, Yuichiro Ueda, Katharina Günther, Constantin Lapa, Mehrbod S. Javadi, Kazuhito Fukushima, Frank Edenhofer, Takahiro Higuchi , Metabolic substrate shift in human induced pluripotent stem cells during cardiac differentiation: Functional assessment using in vitro radionuclide uptake assay. Ijca (2018), doi:10.1016/j.ijcard.2018.06.089. (Preprint)
CC BY-NC-ND: Creative-Commons-Lizenz: Namensnennung, Nicht kommerziell, Keine Bearbeitungen 4.0 International
Naoko Nose
Rudolf A. Werner
Yuichiro Ueda
Katharina Günther
Constantin Lapa
Mehrbod S. Javadi
Kazuhito Fukushima
Frank Edenhofer
Takahiro Higuchi
eng
uncontrolled
tracer
deu
swd
Stammzelle
eng
uncontrolled
induced pluripotent stem cells
eng
uncontrolled
cardiomyocytes
eng
uncontrolled
fatty acid
eng
uncontrolled
stem cell therapy
eng
uncontrolled
hiPSC-CM
Medizin und Gesundheit
open_access
Klinik und Poliklinik für Nuklearmedizin
Institut für Anatomie und Zellbiologie
OpenAIRE
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/16332/Werner_Rudolf_Stammzellen.pdf
17069
2018
eng
229-234
269
article
1
2018-10-30
--
--
Metabolic substrate shift in human induced pluripotent stem cells during cardiac differentiation: Functional assessment using in vitro radionuclide uptake assay
BACKGROUND:
Recent developments in cellular reprogramming technology enable the production of virtually unlimited numbers of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM). Although hiPSC-CM share various characteristic hallmarks with endogenous cardiomyocytes, it remains a question as to what extent metabolic characteristics are equivalent to mature mammalian cardiomyocytes. Here we set out to functionally characterize the metabolic status of hiPSC-CM in vitro by employing a radionuclide tracer uptake assay.
MATERIAL AND METHODS:
Cardiac differentiation of hiPSC was induced using a combination of well-orchestrated extrinsic stimuli such as WNT activation (by CHIR99021) and BMP signalling followed by WNT inhibition and lactate based cardiomyocyte enrichment. For characterization of metabolic substrates, dual tracer uptake studies were performed with \(^{18}\)F‑2‑fluoro‑2‑deoxy‑d‑glucose (\(^{18}\)F-FDG) and \(^{125}\)I‑β‑methyl‑iodophenyl‑pentadecanoic acid (\(^{125}\)I-BMIPP) as transport markers of glucose and fatty acids, respectively.
RESULTS:
After cardiac differentiation of hiPSCs, in vitro tracer uptake assays confirmed metabolic substrate shift from glucose to fatty acids that was comparable to those observed in native isolated human cardiomyocytes. Immunostaining further confirmed expression of fatty acid transport and binding proteins on hiPSC-CM.
CONCLUSIONS:
During in vitro cardiac maturation, we observed a metabolic shift to fatty acids, which are known as a main energy source of mammalian hearts, suggesting hi-PSC-CM as a potential functional phenotype to investigate alteration of cardiac metabolism in cardiac diseases. Results also highlight the use of available clinical nuclear medicine tracers as functional assays in stem cell research for improved generation of autologous differentiated cells for numerous biomedical applications.
International Journal of Cardiology
urn:nbn:de:bvb:20-opus-170699
International Journal of Cardiology 269 (2018) 229–234. DOI: 10.1016/j.ijcard.2018.06.089
701983
Johns Hopkins School of Medicine, The Russell H Morgan Department of Radiology and Radiological Science, Baltimore, MD, USA
Department of Biomedical Imaging, National Cerebral and Cardiovascular Research Center, Suita, Japan
Institut for Molecular Biology and CMBI, Department of Genomics, Stem Cell Biology and Regenerative Medicine, Leopold-Franzens-University Innsbruck, Innsbruck, Austria
Division of Medical Technology and Science, Department of Medical Physics and Engineering, Course of Health Science, Osaka University Graduate School of Medicine, Suita Japan
CC BY-NC-ND: Creative-Commons-Lizenz: Namensnennung, Nicht kommerziell, Keine Bearbeitungen 4.0 International
Naoko Nose
Rudolf A. Werner
Yuichiro Ueda
Katharina Günther
Constantin Lapa
Mehrbod S. Javadi
Kazuhito Fukushima
Frank Edenhofer
Takahiro Higuchi
eng
uncontrolled
tracer
deu
swd
Stammzelle
eng
uncontrolled
induced pluripotent stem cells
eng
uncontrolled
cardiomyocytes
eng
uncontrolled
fatty acid
eng
uncontrolled
stem cell therapy
eng
uncontrolled
hiPSC-CM
Medizin und Gesundheit
open_access
Klinik und Poliklinik für Nuklearmedizin
Institut für Anatomie und Zellbiologie
OpenAIRE
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/17069/Rudolf_Werner_Int_J_Cardiol_Metabolic_substrate_2018.pdf