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Cardiothoracic and Vascular Anesthesia
Anesthesia and Pain Medicine 2013;8(3):175-180.
Published online July 30, 2013.
Isoflurane decreases apoptosis of human embryonic stem cell-derived cardiac progenitor cell under oxidative stress
Bon Wook Koo, Jin Hee Kim, Sung Hee Han, Mi Hyun Kim, Ji Seok Baik
Department of Anesthesiology and Pain Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
Despite the great potential of human embryonic stem cell (hESC)-derived cardiac progenitor cells (CPCs) in the cardiac cell transplantation, the low graft survival still remains as one of the main obstacles in the way to its clinical application. We investigated whether pre-treatment with isoflurane can decrease apoptosis of hESC-derived CPCs under oxidative stress.
Undifferentiated hESCs were differentiated in suspension media with 20% fetal bovine serum (FBS) and 20 ng/ml of bone morphogenetic protein (BMP)-4 through embryoid bodies and grown onto Matrigel-coated plates for 2 or 3 weeks. To identify the differentiated CPCs, immunostaining for nonspecific transcriptional marker (Nkx2.5) was performed. The CPCs were exposed to oxidative stress induced by Fenton reaction with H2O2 and FeSO4. For anesthetic preconditioning, CPCs were exposed to isoflurane (5 vol%) in an isolated chamber. Apoptosis of CPCs was determined by TUNEL staining and detection of activated caspase-3 cell.
hESC-derived CPCs stained with Nkx2.5 were 95 +/- 3% of total cell number. Concentration of isoflurane in the media was 1.1 mM (2.2 MAC). Pretreatment of CPCs with isoflurane showed a significantly lower TUNEL (+) ratio as well as activated caspase-3 cell number compared to control.
Isoflurane decreased hESC-derived Nkx2.5+ CPCs apoptosis induced by oxidative stress. This result suggests that anti-apoptotic effect may play a role in the protective effect of isoflurane.
Key Words: Apoptosis, Caspase-3, Isoflurane, Oxidative stress
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