{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T21:02:32Z","timestamp":1782421352454,"version":"3.54.5"},"reference-count":93,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2024,9,5]],"date-time":"2024-09-05T00:00:00Z","timestamp":1725494400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Brown University 2023 OVPR Seed Award","award":["R01 HL173938"],"award-info":[{"award-number":["R01 HL173938"]}]},{"name":"National Institute of Health","award":["R01 HL173938"],"award-info":[{"award-number":["R01 HL173938"]}]},{"name":"Brown University School of Engineering Carl Nielsen \u201856 Fellowship and Royce Fellowship","award":["R01 HL173938"],"award-info":[{"award-number":["R01 HL173938"]}]},{"name":"NSF-GRFP","award":["R01 HL173938"],"award-info":[{"award-number":["R01 HL173938"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Acute myocardial infarction (MI) is a sudden, severe cardiac ischemic event that results in the death of up to one billion cardiomyocytes (CMs) and subsequent decrease in cardiac function. Engineered cardiac tissues (ECTs) are a promising approach to deliver the necessary mass of CMs to remuscularize the heart. However, the hypoxic environment of the heart post-MI presents a critical challenge for CM engraftment. Here, we present a high-throughput, systematic study targeting several physiological features of human induced pluripotent stem cell-derived CMs (hiPSC-CMs), including metabolism, Wnt signaling, substrate, heat shock, apoptosis, and mitochondrial stabilization, to assess their efficacy in promoting ischemia resistance in hiPSC-CMs. The results of 2D experiments identify hypoxia preconditioning (HPC) and metabolic conditioning as having a significant influence on hiPSC-CM function in normoxia and hypoxia. Within 3D engineered cardiac tissues (ECTs), metabolic conditioning with maturation media (MM), featuring high fatty acid and calcium concentration, results in a 1.5-fold increase in active stress generation as compared to RPMI\/B27 control ECTs in normoxic conditions. Yet, this functional improvement is lost after hypoxia treatment. Interestingly, HPC can partially rescue the function of MM-treated ECTs after hypoxia. Our systematic and iterative approach provides a strong foundation for assessing and leveraging in vitro culture conditions to enhance the hypoxia resistance, and thus the successful clinical translation, of hiPSC-CMs in cardiac regenerative therapies.<\/jats:p>","DOI":"10.3390\/ijms25179627","type":"journal-article","created":{"date-parts":[[2024,9,5]],"date-time":"2024-09-05T06:44:29Z","timestamp":1725518669000},"page":"9627","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Advancing Human iPSC-Derived Cardiomyocyte Hypoxia Resistance for Cardiac Regenerative Therapies through a Systematic Assessment of In Vitro Conditioning"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-1821-3375","authenticated-orcid":false,"given":"Caroline A.","family":"Snyder","sequence":"first","affiliation":[{"name":"Institute for Biology, Engineering and Medicine, School of Engineering, Brown University, Providence, RI 02912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0087-5267","authenticated-orcid":false,"given":"Kiera D.","family":"Dwyer","sequence":"additional","affiliation":[{"name":"Institute for Biology, Engineering and Medicine, School of Engineering, Brown University, Providence, RI 02912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6664-339X","authenticated-orcid":false,"given":"Kareen L. K.","family":"Coulombe","sequence":"additional","affiliation":[{"name":"Institute for Biology, Engineering and Medicine, School of Engineering, Brown University, Providence, RI 02912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,9,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e139","DOI":"10.1161\/CIR.0000000000000757","article-title":"Heart Disease and Stroke Statistics\u20142020 Update: A Report From the American Heart Association","volume":"141","author":"Virani","year":"2020","journal-title":"Circulation"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1038\/nbt1117","article-title":"Regenerating the heart","volume":"23","author":"Laflamme","year":"2005","journal-title":"Nat. Biotechnol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1161\/CIRCULATIONAHA.117.029343","article-title":"Cardiomyocyte regeneration: A consensus statement","volume":"136","author":"Eschenhagen","year":"2017","journal-title":"Circulation"},{"key":"ref_4","first-page":"222","article-title":"Heart failure after myocardial infarction: Incidence and predictors","volume":"8","author":"Stehlik","year":"2020","journal-title":"ESC Heart Fail."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1991","DOI":"10.1161\/CIRCULATIONAHA.120.047904","article-title":"Human Engineered Heart Tissue Patches Remuscularize the Injured Heart in a Dose-Dependent Manner","volume":"143","author":"Querdel","year":"2021","journal-title":"Circulation"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.yjmcc.2021.10.005","article-title":"How to repair a broken heart with pluripotent stem cell-derived cardiomyocytes","volume":"163","author":"Eschenhagen","year":"2021","journal-title":"J. Mol. Cell. Cardiol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Dwyer, K.D., Kant, R.J., Soepriatna, A.H., Roser, S.M., Daley, M.C., Sabe, S.A., Xu, C.M., Choi, B.-R., Sellke, F.W., and Coulombe, K.L. (2023). One billion hiPSC-cardiomyocytes: Upscaling engineered cardiac tissues to create high cell density therapies for clinical translation in heart regeneration. Bioengineering, 10.","DOI":"10.3390\/bioengineering10050587"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Khalil, N.N., and McCain, M.L. (2021). Engineering the Cellular Microenvironment of Post-infarct Myocardium on a Chip. Front. Cardiovasc. Med., 8.","DOI":"10.3389\/fcvm.2021.709871"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Handley, E., and Callanan, A. (2022). Modulation of Tissue Microenvironment Following Myocardial Infarction. Adv. NanoBiomed Res., 2.","DOI":"10.1002\/anbr.202200005"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Yang, G., Mahadik, B., Choi, J.Y., and Fisher, J.P. (2020). Vascularization in tissue engineering: Fundamentals and state-of-art. Prog. Biomed. Eng., 2.","DOI":"10.1088\/2516-1091\/ab5637"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"342ps13","DOI":"10.1126\/scitranslmed.aad2304","article-title":"Distilling complexity to advance cardiac tissue engineering","volume":"8","author":"Ogle","year":"2016","journal-title":"Sci. Transl. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1038\/s41569-019-0331-x","article-title":"Cardiomyocyte maturation: Advances in knowledge and implications for regenerative medicine","volume":"17","author":"Karbassi","year":"2020","journal-title":"Nat. Rev. Cardiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"107925","DOI":"10.1016\/j.celrep.2020.107925","article-title":"Metabolic maturation media improve physiological function of human iPSC-derived cardiomyocytes","volume":"32","author":"Feyen","year":"2020","journal-title":"Cell Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1557","DOI":"10.1161\/CIRCULATIONAHA.114.014998","article-title":"Mechanical Stress Conditioning and Electrical Stimulation Promote Contractility and Force Maturation of Induced Pluripotent Stem Cell-Derived Human Cardiac Tissue","volume":"134","author":"Ruan","year":"2016","journal-title":"Circulation"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1412","DOI":"10.1161\/CIRCULATIONAHA.121.053563","article-title":"In Vitro Matured Human Pluripotent Stem Cell\u2013Derived Cardiomyocytes Form Grafts With Enhanced Structure and Function in Injured Hearts","volume":"145","author":"Dhahri","year":"2022","journal-title":"Circulation"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1161\/01.CIR.100.2.193","article-title":"Survival, integration, and differentiation of cardiomyocyte grafts: A study in normal and injured rat hearts","volume":"100","author":"Reinecke","year":"1999","journal-title":"Circulation"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1093\/stcltm\/szac061","article-title":"Metabolic maturation increases susceptibility to hypoxia-induced damage in human iPSC-derived cardiomyocytes","volume":"11","author":"Peters","year":"2022","journal-title":"Stem Cells Transl. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.stem.2020.06.001","article-title":"Wnt activation and reduced cell-cell contact synergistically induce massive expansion of functional human iPSC-derived cardiomyocytes","volume":"27","author":"Buikema","year":"2020","journal-title":"Cell Stem Cell."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.freeradbiomed.2022.08.005","article-title":"Inhibition of mitochondrial superoxide promotes the development of hiPS-CMs during differentiation","volume":"190","author":"Li","year":"2022","journal-title":"Free. Radic. Biol. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1038\/sj.bjp.0704908","article-title":"Cyclosporine A regulate oxidative stress-induced apoptosis in cardiomyocytes: Mechanisms via ROS generation, iNOS and Hsp70","volume":"137","author":"Chen","year":"2002","journal-title":"Br. J. Pharmacology."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2034","DOI":"10.1021\/acs.molpharmaceut.0c00192","article-title":"Targeted Delivery of Recombinant Heat Shock Protein 27 to Cardiomyocytes Promotes Recovery from Myocardial Infarction","volume":"17","author":"Kim","year":"2020","journal-title":"Mol. Pharm."},{"key":"ref_22","first-page":"291","article-title":"Heat Shock Proteins: Protection and Potential Biomarkers for Ischemic Injury of Cardiomyocytes After Surgery","volume":"33","author":"Lollo","year":"2018","journal-title":"Rev. Bras. Cir. Cardiovasc."},{"key":"ref_23","first-page":"2617","article-title":"Moderate hypothermia induces protection against hypoxia\/reoxygenation injury by enhancing SUMOylation in cardiomyocytes","volume":"22","author":"Chen","year":"2020","journal-title":"Mol. Med. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e026160","DOI":"10.1161\/JAHA.122.026160","article-title":"Therapeutic Hypothermia Inhibits Hypoxia-Induced Cardiomyocyte Apoptosis Via the MiR-483-3p\/Cdk9 Axis","volume":"12","author":"Xue","year":"2023","journal-title":"J. Am. Heart Assoc."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.yexmp.2015.11.025","article-title":"Hypoxic preconditioning protects cardiomyocytes against hypoxia\/reoxygenation-induced cell apoptosis via sphingosine kinase 2 and FAK\/AKT pathway","volume":"100","author":"Zhang","year":"2015","journal-title":"Exp. Mol. Pathol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1161\/01.RES.86.5.541","article-title":"Preconditioning in Cardiomyocytes Protects by Attenuating Oxidant Stress at Reperfusion","volume":"86","author":"Hoek","year":"2000","journal-title":"Circ. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1016\/j.stemcr.2019.08.013","article-title":"Fatty Acids Enhance the Maturation of Cardiomyocytes Derived from Human Pluripotent Stem Cells","volume":"13","author":"Yang","year":"2019","journal-title":"Stem Cell Rep."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1161\/CIRCRESAHA.118.313249","article-title":"Metabolic maturation of human pluripotent stem cell-derived cardiomyocytes by inhibition of HIF1\u03b1 and LDHA","volume":"123","author":"Hu","year":"2018","journal-title":"Circ. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1016\/j.stemcr.2018.01.039","article-title":"Contractile work contributes to maturation of energy metabolism in hiPSC-derived cardiomyocytes","volume":"10","author":"Ulmer","year":"2018","journal-title":"Stem Cell Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"E8372","DOI":"10.1073\/pnas.1707316114","article-title":"Functional screening in human cardiac organoids reveals a metabolic mechanism for cardiomyocyte cell cycle arrest","volume":"114","author":"Mills","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3479","DOI":"10.1529\/biophysj.107.124545","article-title":"Substrate Stiffness Affects the Functional Maturation of Neonatal Rat Ventricular Myocytes","volume":"95","author":"Jacot","year":"2008","journal-title":"Biophys. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1567","DOI":"10.1039\/C7BM00323D","article-title":"Anisotropic microfibrous scaffolds enhance the organization and function of cardiomyocytes derived from induced pluripotent stem cells","volume":"5","author":"Wanjare","year":"2017","journal-title":"Biomater. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1643","DOI":"10.1177\/1535370217701006","article-title":"Toward improved myocardial maturity in an organ-on-chip platform with immature cardiac myocytes","volume":"242","author":"Sheehy","year":"2017","journal-title":"Exp. Biol. Med."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1038\/s41586-018-0016-3","article-title":"Advanced maturation of human cardiac tissue grown from pluripotent stem cells","volume":"556","author":"Ma","year":"2018","journal-title":"Nature"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2633","DOI":"10.1172\/JCI26471","article-title":"Death versus survival: Functional interaction between the apoptotic and stress-inducible heat shock protein pathways","volume":"115","author":"Beere","year":"2005","journal-title":"J. Clin. Investig."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"H216","DOI":"10.1152\/ajpheart.00060.2018","article-title":"Cardioprotection by intermittent hypoxia conditioning: Evidence, mechanisms, and therapeutic potential","volume":"315","author":"Mallet","year":"2018","journal-title":"Am. J. Physiol. Circ. Physiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1016\/j.acthis.2013.01.005","article-title":"Experimental acute myocardial infarction in rats: HIF-1\u03b1, caspase-3, erythropoietin and erythropoietin receptor expression and the cardioprotective effects of two different erythropoietin doses","volume":"115","author":"Bagla","year":"2013","journal-title":"Acta Histochem."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Pescador, N., Villar, D., Cifuentes, D., Garcia-Rocha, M., Ortiz-Barahona, A., Vazquez, S., Ordo\u00f1ez, A., Cuevas, Y., Saez-Morales, D., and Garcia-Bermejo, M.L. (2010). Hypoxia Promotes Glycogen Accumulation through Hypoxia Inducible Factor (HIF)-Mediated Induction of Glycogen Synthase 1. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0009644"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1007\/s00395-005-0537-4","article-title":"Erythropoietin protects the myocardium against reperfusion injury in vitro and in vivo","volume":"100","author":"Bullard","year":"2005","journal-title":"Basic Res. Cardiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"12567","DOI":"10.1074\/jbc.274.18.12567","article-title":"Glucose Uptake and Glycolysis Reduce Hypoxia-induced Apoptosis in Cultured Neonatal Rat Cardiac Myocytes","volume":"274","author":"Malhotra","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Kant, R.J., Dwyer, K.D., Lee, J.-H., Polucha, C., Kobayashi, M., Pyon, S., Soepriatna, A.H., Lee, J., and Coulombe, K.L.K. (2023). Patterned Arteriole-Scale Vessels Enhance Engraftment, Perfusion, and Vessel Branching Hierarchy of Engineered Human Myocardium for Heart Regeneration. Cells, 12.","DOI":"10.3390\/cells12131698"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"9363809","DOI":"10.1155\/2020\/9363809","article-title":"Human Cardiac Fibroblast Number and Activation State Modulate Electromechanical Function of hiPSC-Cardiomyocytes in Engineered Myocardium","volume":"2020","author":"Rupert","year":"2020","journal-title":"Stem Cells International."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.semcdb.2021.05.022","article-title":"Maturation of human pluripotent stem cell derived cardiomyocytes in vitro and in vivo","volume":"118","author":"Quesnel","year":"2021","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Bian, W., Jackman, C.P., and Bursac, N. (2014). Controlling the structural and functional anisotropy of engineered cardiac tissues. Biofabrication, 6.","DOI":"10.1088\/1758-5082\/6\/2\/024109"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"237","DOI":"10.5301\/ijao.5000084","article-title":"Engineering of Oriented Myocardium on Three-Dimensional Micropatterned Collagen-Chitosan Hydrogel","volume":"35","author":"Chiu","year":"2012","journal-title":"Int. J. Artif. Organs"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1991","DOI":"10.1089\/scd.2012.0490","article-title":"Structural and Functional Maturation of Cardiomyocytes Derived from Human Pluripotent Stem Cells","volume":"22","author":"Lundy","year":"2013","journal-title":"Stem Cells Dev."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.scr.2017.10.011","article-title":"Substrate and mechanotransduction influence SERCA2a localization in human pluripotent stem cell-derived cardiomyocytes affecting functional performance","volume":"25","author":"Martewicz","year":"2017","journal-title":"Stem Cell Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"586","DOI":"10.1016\/j.stem.2018.09.009","article-title":"Single-Cell Transcriptomic Analysis of Cardiac Differentiation from Human PSCs Reveals HOPX-Dependent Cardiomyocyte Maturation","volume":"23","author":"Friedman","year":"2018","journal-title":"Cell Stem Cell"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1397","DOI":"10.1089\/scd.2016.0073","article-title":"From Early Embryonic to Adult Stage: Comparative Study of Action Potentials of Native and Pluripotent Stem Cell-Derived Cardiomyocytes","volume":"25","author":"Peinkofer","year":"2016","journal-title":"Stem Cells Dev."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"e13862","DOI":"10.14814\/phy2.13862","article-title":"Pacemaking in cardiac tissue. From IK2 to a coupled-clock system","volume":"7","author":"Carmeliet","year":"2019","journal-title":"Physiol. Rep."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"e005135","DOI":"10.1161\/JAHA.116.005135","article-title":"Switch From Fetal to Adult SCN5A Isoform in Human Induced Pluripotent Stem Cell\u2013Derived Cardiomyocytes Unmasks the Cellular Phenotype of a Conduction Disease\u2013Causing Mutation","volume":"6","author":"Veerman","year":"2017","journal-title":"J. Am. Heart Assoc."},{"key":"ref_52","first-page":"103","article-title":"Beyond pharmaceuticals: Fit-for-purpose new approach methodologies for environmental cardiotoxicity testing","volume":"40","author":"Daley","year":"2022","journal-title":"Altex"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1161\/CIRCRESAHA.111.250266","article-title":"Induced Pluripotent Stem Cells in Cardiovascular Drug Discovery","volume":"112","author":"Mercola","year":"2013","journal-title":"Circ. Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1016\/j.stemcr.2019.04.005","article-title":"Human Embryonic Stem Cell-Derived Cardiomyocytes Regenerate the Infarcted Pig Heart but Induce Ventricular Tachyarrhythmias","volume":"12","author":"Romagnuolo","year":"2019","journal-title":"Stem Cell Rep."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1038\/nature13233","article-title":"Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts","volume":"510","author":"Chong","year":"2014","journal-title":"Nature"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1038\/nbt.4162","article-title":"Human embryonic stem cell\u2013derived cardiomyocytes restore function in infarcted hearts of non-human primates","volume":"36","author":"Liu","year":"2018","journal-title":"Nat. Biotechnol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"dmm049834","DOI":"10.1242\/dmm.049834","article-title":"Immature human engineered heart tissues engraft in a guinea pig chronic injury model","volume":"16","author":"Shibamiya","year":"2023","journal-title":"Dis. Model. Mech."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1038\/nature11317","article-title":"Human ES-cell-derived cardiomyocytes electrically couple and suppress arrhythmias in injured hearts","volume":"489","author":"Shiba","year":"2012","journal-title":"Nature"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.semcdb.2021.04.019","article-title":"Maturing heart muscle cells: Mechanisms and transcriptomic insights","volume":"119","author":"Murphy","year":"2021","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"26","DOI":"10.14797\/mdcvj.1309","article-title":"Metabolic Control of Cardiomyocyte Cell Cycle","volume":"19","author":"Garry","year":"2023","journal-title":"Methodist DeBakey Cardiovasc. J."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/0006-8993(89)90144-3","article-title":"Survival and growth of hippocampal neurons in defined medium at low density: Advantages of a sandwich culture technique or low oxygen","volume":"494","author":"Brewer","year":"1989","journal-title":"Brain Res."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"H224","DOI":"10.1152\/ajpheart.00689.2006","article-title":"Role of oxidative stress in PKC-\u03b4 upregulation and cardioprotection induced by chronic intermittent hypoxia","volume":"292","year":"2007","journal-title":"Am. J. Physiol. Circ. Physiol."},{"key":"ref_63","first-page":"48","article-title":"Effect of adaptation to continuous and intermittent hypoxia on heart resistance to ischemic and reperfusion arrhythmias","volume":"3","author":"Meerson","year":"1989","journal-title":"Patol. Fiziol. I Eksperimental\u2019naia Ter."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"H1566","DOI":"10.1152\/ajpheart.00586.2004","article-title":"Increased expression and altered subcellular distribution of PKC-\u03b4 in chronically hypoxic rat myocardium: Involvement in cardioprotection","volume":"288","year":"2005","journal-title":"Am. J. Physiol. Circ. Physiol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"H1695","DOI":"10.1152\/ajpheart.00276.2011","article-title":"Intermittent hypobaric hypoxia improves postischemic recovery of myocardial contractile function via redox signaling during early reperfusion","volume":"301","author":"Wang","year":"2011","journal-title":"Am. J. Physiol. Circ. Physiol."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Milano, G., Abruzzo, P.M., Bolotta, A., Marini, M., Terraneo, L., Ravara, B., Gorza, L., Vitadello, M., Burattini, S., and Curzi, D. (2013). Impact of the Phosphatidylinositide 3-Kinase Signaling Pathway on the Cardioprotection Induced by Intermittent Hypoxia. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0076659"},{"key":"ref_67","first-page":"597","article-title":"Effects of adaptation to intermittent high altitude hypoxia on ischemic ventricular arrhythmias in rats","volume":"49","author":"Asemu","year":"2000","journal-title":"Physiol. Res."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1007\/s12576-015-0376-5","article-title":"K ATP channels and MPTP are involved in the cardioprotection bestowed by chronic intermittent hypobaric hypoxia in the developing rat","volume":"65","author":"Bu","year":"2015","journal-title":"J. Physiol. Sci."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1016\/S0008-6363(02)00456-X","article-title":"Effects of mitochondrial KATP modulators on cardioprotection induced by chronic high altitude hypoxia in rats","volume":"55","author":"Koten","year":"2002","journal-title":"Cardiovasc. Res."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1113\/expphysiol.2012.065102","article-title":"Mitochondrial energy metabolism plays a critical role in the cardioprotection afforded by intermittent hypobaric hypoxia","volume":"97","author":"Wang","year":"2012","journal-title":"Exp. Physiol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1161\/01.RES.80.5.699","article-title":"Myocyte Adaptation to Chronic Hypoxia and Development of Tolerance to Subsequent Acute Severe Hypoxia","volume":"80","author":"Silverman","year":"1997","journal-title":"Circ. Res."},{"key":"ref_72","first-page":"463","article-title":"Hypoxic preconditioning induces endoplasmic reticulum stress-related cardioprotection mediated by p38 mitogen-activated protein kinase","volume":"58","author":"Zhu","year":"2006","journal-title":"Sheng Li Xue Bao"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"C2046","DOI":"10.1152\/ajpcell.00458.2006","article-title":"Chronic intermittent hypoxia alters Ca2+ handling in rat cardiomyocytes by augmented Na+\/Ca2+ exchange and ryanodine receptor activities in ischemia-reperfusion","volume":"292","author":"Yeung","year":"2007","journal-title":"Am. J. Physiol.-Cell Physiol."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"H2280","DOI":"10.1152\/ajpheart.01164.2010","article-title":"Enhancement of Na\/K pump activity by chronic intermittent hypobaric hypoxia protected against reperfusion injury","volume":"300","author":"Guo","year":"2011","journal-title":"Am. J. Physiol. Circ. Physiol."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1016\/j.yjmcc.2005.09.016","article-title":"Intermittent high altitude hypoxia inhibits opening of mitochondrial permeability transition pores against reperfusion injury","volume":"40","author":"Zhu","year":"2006","journal-title":"J. Mol. Cell. Cardiol."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/j.xjon.2023.05.013","article-title":"Intramyocardial injection of hypoxia-conditioned extracellular vesicles modulates apoptotic signaling in chronically ischemic myocardium","volume":"15","author":"Harris","year":"2023","journal-title":"JTCVS Open"},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Sabe, S.A., Xu, C.M., Potz, B.A., Malhotra, A., Sabra, M., Harris, D.D., Broadwin, M., Abid, M.R., and Sellke, F.W. (2023). Comparative Analysis of Normoxia- and Hypoxia-Modified Extracellular Vesicle Therapy in Function, Perfusion, and Collateralization in Chronically Ischemic Myocardium. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24032076"},{"key":"ref_78","doi-asserted-by":"crossref","unstructured":"Xu, C.M., Karbasiafshar, C., Teixeira, R.B., Ahsan, N., Corssac, G.B., Sellke, F.W., and Abid, M.R. (2023). Proteomic Assessment of Hypoxia-Pre-Conditioned Human Bone Marrow Mesenchymal Stem Cell-Derived Extracellular Vesicles Demonstrates Promise in the Treatment of Cardiovascular Disease. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24021674"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1186\/s12964-024-01722-7","article-title":"Hypoxic extracellular vesicles from hiPSCs protect cardiomyocytes from oxidative damage by transferring antioxidant proteins and enhancing Akt\/Erk\/NRF2 signaling","volume":"22","author":"Paw","year":"2024","journal-title":"Cell Commun. Signal."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1007\/s00395-016-0538-5","article-title":"\u03b4-Opioid receptor (DOR) signaling and reactive oxygen species (ROS) mediate intermittent hypoxia induced protection of canine myocardium","volume":"111","author":"Estrada","year":"2016","journal-title":"Basic Res. Cardiol."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1007\/s00395-006-0599-y","article-title":"\u03b21-Adrenergic receptor antagonism abrogates cardioprotective effects of intermittent hypoxia","volume":"101","author":"Mallet","year":"2006","journal-title":"Basic Res. Cardiol."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"806","DOI":"10.1177\/153537020422900813","article-title":"Intermittent Hypoxic Training Protects Canine Myocardium from Infarction","volume":"229","author":"Zong","year":"2004","journal-title":"Exp. Biol. Med."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"10986","DOI":"10.3892\/etm.2021.10986","article-title":"Targeting angiogenesis in myocardial infarction: Novel therapeutics (Review)","volume":"23","author":"Li","year":"2021","journal-title":"Exp. Ther. Med."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.actbio.2018.01.017","article-title":"Integrated approaches to spatiotemporally directing angiogenesis in host and engineered tissues","volume":"69","author":"Kant","year":"2018","journal-title":"Acta Biomater."},{"key":"ref_85","doi-asserted-by":"crossref","unstructured":"Chen, C., Wang, J., Liu, C., Hu, J., and Liu, L. (2023). Pioneering therapies for post-infarction angiogenesis: Insight into molecular mechanisms and preclinical studies. Biomed. Pharmacother., 166.","DOI":"10.1016\/j.biopha.2023.115306"},{"key":"ref_86","doi-asserted-by":"crossref","unstructured":"Munarin, F., Kant, R.J., Rupert, C.E., Khoo, A., and Coulombe, K.L. (2020). Engineered human myocardium with local release of angiogenic proteins improves vascularization and cardiac function in injured rat hearts. Biomater., 251.","DOI":"10.1016\/j.biomaterials.2020.120033"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"S145","DOI":"10.1161\/CIRCULATIONAHA.107.757286","article-title":"Endothelial Cell Coculture Within Tissue-Engineered Cardiomyocyte Sheets Enhances Neovascularization and Improves Cardiac Function of Ischemic Hearts","volume":"118","author":"Sekine","year":"2008","journal-title":"Circulation"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1089\/ten.tea.2009.0130","article-title":"Transplantation of a Tissue-Engineered Human Vascularized Cardiac Muscle","volume":"16","author":"Lesman","year":"2010","journal-title":"Tissue Eng. Part A"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"16568","DOI":"10.1073\/pnas.0908381106","article-title":"Physiological function and transplantation of scaffold-free and vascularized human cardiac muscle tissue","volume":"106","author":"Stevens","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1038\/nprot.2012.150","article-title":"Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt\/\u03b2-catenin signaling under fully defined conditions","volume":"8","author":"Lian","year":"2012","journal-title":"Nat. Protoc."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1038\/nmeth.2999","article-title":"Chemically defined generation of human cardiomyocytes","volume":"11","author":"Burridge","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"2040","DOI":"10.1021\/acsbiomaterials.2c00162","article-title":"Optimization of a PDMS-Based Cell Culture Substrate for High-Density Human-Induced Pluripotent Stem Cell Adhesion and Long-Term Differentiation into Cardiomyocytes under a Xeno-Free Condition","volume":"8","author":"Etezadi","year":"2022","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1089\/ten.tec.2017.0068","article-title":"Laser-Etched Designs for Molding Hydrogel-Based Engineered Tissues","volume":"23","author":"Munarin","year":"2017","journal-title":"Tissue Eng. 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