Supported by grant (04-2008-007) from the SNUBH Research Fund and grant (SC1150) from Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology, Korea.
Isoflurane decreases death of human embryonic stem cell-derived, transcriptional marker Nkx2.5+ cardiac progenitor cells
Article first published online: 7 SEP 2011
© 2011 The Authors Acta Anaesthesiologica Scandinavica © 2011 The Acta Anaesthesiologica Scandinavica Foundation
Acta Anaesthesiologica Scandinavica
Volume 55, Issue 9, pages 1124–1131, October 2011
How to Cite
KIM, J. H., OH, A. Y., CHOI, Y. M., KU, S. Y., KIM, Y. Y., LEE, N. J., SEPAC, A. and BOSNJAK, Z. J. (2011), Isoflurane decreases death of human embryonic stem cell-derived, transcriptional marker Nkx2.5+ cardiac progenitor cells. Acta Anaesthesiologica Scandinavica, 55: 1124–1131. doi: 10.1111/j.1399-6576.2011.02509.x
- Issue published online: 21 SEP 2011
- Article first published online: 7 SEP 2011
- Manuscript Accepted: 9 JUL 2011
- SNUBH Research Fund. Grant Number: 04-2008-007
- Stem Cell Research Center of the 21st Century Frontier Research Program. Grant Number: SC1150
Cardiac progenitor cells (CPCs) derived from human embryonic stem cells (hESCs) can multiply and generate cardiomyocytes, offering their tremendous potential for cardiac regenerative therapy. However, poor survival under stressful conditions is a major hurdle in the regeneration. We investigated whether isoflurane-induced preconditioning can increase hESC-derived CPC survival under oxidative stress.
Undifferentiated hESCs were cultured in suspension with 20% FBS (fetal bovine serum) and 20 ng/ml of BMP-4 (bone morphogenetic protein-4) to form embryoid bodies and grown onto Matrigel-coated plates for 2–3 weeks. To characterise the differentiated CPCs, immunostaining for Nkx2.5 (nonspecific transcriptional marker) and Isl-1 was performed. hESC-derived CPCs were exposed to oxidative stress induced by H2O2 and FeSO4. For anaesthetic preconditioning, CPCs were exposed to isoflurane (0.25, 0.5, 1.0 mM). CPC survival was determined by trypan blue exclusion. A mitoKATP channels inhibitor, 5-hydroxydecanoic acid (200 μM) and an opener, diazoxide (100 μM), were used to investigate the involvement of mitoKATP channels.
hESC-derived CPCs stained with Nkx2.5 were 95 ± 3% of total cell number. Isoflurane (0.5 and 1.0 mM)-preconditioned CPCs showed a significantly lower death rate compared with control (0.5 mM: 30.6 ± 10.7% and 1.0 mM: 28.5 ± 6.2% vs. control: 43.2 ± 9.9%). Inhibition of mitoKATP channels with 5-HD completely abolished the protective effects of isoflurane. Diazoxide significantly decreased CPC death (29.5 ± 12.4%). However, when diazoxide was applied to CPC preconditioned with isoflurane, CPC death did not decrease further (28.7 ± 10.9%).
Isoflurane increased hESC-derived Nkx2.5+CPC survival under oxidative stress, and mitoKATP channels may be involved in the protective effect.