Hypoxic preconditioning induces the expression of prosurvival and proangiogenic markers in mesenchymal stem cells.
Stem cells transplanted to the ischemic myocardium usually encounter massive cell death within a few days of therapy. Hypoxic preconditioning (HPC) is currently employed as a strategy to prepare stem cells for increased survival and engraftment in the heart. However, HPC of stem cells has provided varying results, supposedly due to the differences in the oxygen concentration, duration of exposure, and passage conditions. In the present study, we determined the effect of HPC on rat mesenchymal stem cells (MSCs) exposed to 0.5% oxygen concentration for 24, 48, or 72 h. We evaluated the expression of prosurvival, proangiogenic, and functional markers such as hypoxia-inducible factor-1α, VEGF, phosphorylated Akt, survivin, p21, cytochrome c, caspase-3, caspase-7, CXCR4, and c-Met. MSCs exposed to 24-h hypoxia showed reduced apoptosis on being subjected to severe hypoxic conditions. They also had significantly higher levels of prosurvival, proangiogenic, and prodifferentiation proteins when compared with longer exposure (72 h). Cells taken directly from the cryopreserved state did not respond effectively to the 24-h HPC as those that were cultured under normoxia before HPC. Cells cultured under normoxia before HPC showed decreased apoptosis, enhanced expression of connexin-43, cardiac myosin heavy chain, and CD31. The preconditioned cells were able to differentiate into the cardiovascular lineage. The results suggest that MSCs cultured under normoxia before 24-h HPC are in a state of optimal expression of prosurvival, proangiogenic, and functional proteins that may increase the survival and engraftment in the infarct heart. These results could provide further insights into optimal preparation of MSCs which would greatly influence the effectiveness of cell therapy in vivo.
Chacko
Simi M
SM
Davis Heart and Lung Research Institute, Division of Cardiovascular Medicine, Department of Internal Medicine, The Ohio State University, Columbus, Ohio 43210, USA.
Ahmed
Shabnam
S
Selvendiran
Karuppaiyah
K
Kuppusamy
M Lakshmi
ML
Khan
Mahmood
M
Kuppusamy
Periannan
P
D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000032:analysis; D053179:Caspase 7 / Q000032:analysis; D015687:Cell Hypoxia; D002460:Cell Line; D018031:Connexin 43 / Q000032:analysis; D045304:Cytochromes c / Q000032:analysis; D006321:Heart; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000032:analysis; D019157:Ischemic Preconditioning, Myocardial; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000032:analysis; D018995:Myosin Heavy Chains / Q000032:analysis; D018919:Neovascularization, Physiologic; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D051057:Proto-Oncogene Proteins c-akt / Q000032:analysis; D019859:Proto-Oncogene Proteins c-met; D051381:Rats; D019718:Receptors, CXCR4 / Q000032:analysis; D000077022:Survivin; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis; D020741:rho GTP-Binding Proteins / Q000032:analysis
C516989:Birc5 protein, rat; D018031:Connexin 43; C486264:Cxcr4 protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008869:Microtubule-Associated Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D019718:Receptors, CXCR4; D000077022:Survivin; D042461:Vascular Endothelial Growth Factor A; D045304:Cytochromes c; D019859:Proto-Oncogene Proteins c-met; D051057:Proto-Oncogene Proteins c-akt; D053148:Caspase 3; D053179:Caspase 7; D018995:Myosin Heavy Chains; D020741:rho GTP-Binding Proteins