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Mol. Cells 2012; 34(2): 149-158

Published online August 31, 2012

https://doi.org/10.1007/s10059-012-0019-0

© The Korean Society for Molecular and Cellular Biology

Liver Cell Line Derived Conditioned Medium Enhances Myofibril Organization of Primary Rat Cardiomyocytes

Jinseok Kim1,2,3,7, Yu-Shik Hwang1,2,4,7, Alice Mira Chung1,2, Bong Geun Chung1,2,5, and Ali Khademhosseini1,2,4,6,*

1Center for Biomedical Engineering, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Cambridge, MA, 02139, USA, 2Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA, 3Center for Bionics, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul 136-791, Korea, 4Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul 130-701, Korea, 5Department of Bionano Engineering, Hanyang University, Ansan 426-791, Korea, 6Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA 02115, USA, 7These authors contributed equally to this work.

Correspondence to : *Correspondence: alik@rics.bwh.harvard.edu

Received: January 13, 2012; Revised: May 29, 2012; Accepted: June 21, 2012

Abstract

Cardiomyocytes are the fundamental cells of the heart and play an important role in engineering of tissue constructs for regenerative medicine and drug discovery. Therefore, the development of culture conditions that can be used to generate functional cardiomyocytes to form cardiac tissue may be of great interest. In this study, isolated neonatal rat cardiomyocytes were cultured with several culture conditions in vitro and characterized for cell proliferation, myofibril organization, and cardiac functionality by assessing cell morphology, immunocytochemical staining, and time-lapse confocal scanning microscopy. When cardiomyocytes were cultured in liver cell line derived conditioned medium without exogenous growth factors and cytokines, the cell proliferation increased, cell morphology was highly elongated, and subsequent myofibril organization was highly developed. These developed myofibril organi-zation also showed high level of contractibility and synchronization, representing high functionality of cardiomyocytes. Interestingly, many of the known factors in hepatic conditioned medium, such as insulin-like growth factor II (IGFII), macrophage colony-stimulating factor (MCSF), leukemia inhibitory factor (LIF), did not show similar effects as the hepatic conditioned medium, suggesting the possibility of synergistic activity of the several soluble factors or the presence of unknown factors in hepatic conditioned medium. Finally, we demonstrated that our culture system could provide a potentially powerful tool for in vitro cardiac tissue organization and cardiac function study.

Keywords cardiomyocytes, hepatic conditioned Medium, myofibril organization, proliferation

Article

Research Article

Mol. Cells 2012; 34(2): 149-158

Published online August 31, 2012 https://doi.org/10.1007/s10059-012-0019-0

Copyright © The Korean Society for Molecular and Cellular Biology.

Liver Cell Line Derived Conditioned Medium Enhances Myofibril Organization of Primary Rat Cardiomyocytes

Jinseok Kim1,2,3,7, Yu-Shik Hwang1,2,4,7, Alice Mira Chung1,2, Bong Geun Chung1,2,5, and Ali Khademhosseini1,2,4,6,*

1Center for Biomedical Engineering, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Cambridge, MA, 02139, USA, 2Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA, 3Center for Bionics, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul 136-791, Korea, 4Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul 130-701, Korea, 5Department of Bionano Engineering, Hanyang University, Ansan 426-791, Korea, 6Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA 02115, USA, 7These authors contributed equally to this work.

Correspondence to:*Correspondence: alik@rics.bwh.harvard.edu

Received: January 13, 2012; Revised: May 29, 2012; Accepted: June 21, 2012

Abstract

Cardiomyocytes are the fundamental cells of the heart and play an important role in engineering of tissue constructs for regenerative medicine and drug discovery. Therefore, the development of culture conditions that can be used to generate functional cardiomyocytes to form cardiac tissue may be of great interest. In this study, isolated neonatal rat cardiomyocytes were cultured with several culture conditions in vitro and characterized for cell proliferation, myofibril organization, and cardiac functionality by assessing cell morphology, immunocytochemical staining, and time-lapse confocal scanning microscopy. When cardiomyocytes were cultured in liver cell line derived conditioned medium without exogenous growth factors and cytokines, the cell proliferation increased, cell morphology was highly elongated, and subsequent myofibril organization was highly developed. These developed myofibril organi-zation also showed high level of contractibility and synchronization, representing high functionality of cardiomyocytes. Interestingly, many of the known factors in hepatic conditioned medium, such as insulin-like growth factor II (IGFII), macrophage colony-stimulating factor (MCSF), leukemia inhibitory factor (LIF), did not show similar effects as the hepatic conditioned medium, suggesting the possibility of synergistic activity of the several soluble factors or the presence of unknown factors in hepatic conditioned medium. Finally, we demonstrated that our culture system could provide a potentially powerful tool for in vitro cardiac tissue organization and cardiac function study.

Keywords: cardiomyocytes, hepatic conditioned Medium, myofibril organization, proliferation

Mol. Cells
Dec 31, 2021 Vol.44 No.12, pp. 861~919
COVER PICTURE
Structure of the fly peripheral neurons in the fly head. Flies have basic sensory organs including eyes for vision, antennae and maxillary palps for olfaction, and proboscis (magenta) for gustation which can be labelled with monoclonal antibody 22C10. The figure is a 3D reconstructed image with 30 slices of confocal sections with 3 μm interval. It shows that the proboscis is required for sensing attractive carboxylic acids such as glycolic acid, citric acid, and lactic acid (Shrestha and Lee, pp. 900-910).

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