Bogoslovsky T
Center for Neuroscience & Regenerative Medicine (CNRM),
(USUHS), 10 Center DR RM 5B06, Bethesda MD 20892
Tanzania
Research Article
Cryopreservation and Enumeration of Human Endothelial Progenitor and Endothelial Cells for Clinical Trials
Author(s): Bogoslovsky T, Wang D, Maric D, Scattergood-Keepper L, Spatz M, Auh S and Hallenbeck J
Bogoslovsky T, Wang D, Maric D, Scattergood-Keepper L, Spatz M, Auh S and Hallenbeck J
Background: Endothelial progenitor cells (EPC) are markers of endothelial injury and may serve as a surrogate marker for vascular repair in interventional clinical trials. Objectives of this study were to modify a method of isolation of peripheral blood mononuclear cells (PBMC) and enumeration of EPC and mature endothelial cells (EC) from peripheral blood and to evaluate influence of cryopreservation on viability of PBMC and on numbers of EPC and EC. Patients/Methods: EPC and EC were analyzed in healthy volunteers in freshly isolated PBMC collected in CPT (cell preparation tubes) and in PBMC cryopreserved with: 1) Gibco RecoveryTM Cell Culture Freezing Medium, 2) custom freezing medium. Viability of PBMC was tested using DAPI. EPC were gated for CD45- CD34+CD133+/-VEGFR2+/- and EC were gated for CD45-CD146+CD34+/-VEGFR2+/-. Results: Cryopreservation for 7 days at -80°C decreased v.. View More»
DOI:
10.4172/2155-9864.1000158