circulating angiogenic cells can be derived from cryopreserved peripheral blood mononuclear cells循环血管生成细胞可以从低温贮藏外周血单核细胞.pdfVIP

Circulating Angiogenic Cells can be Derived from Cryopreserved Peripheral Blood Mononuclear Cells 1,2 1,3 1 1,2 4 Tanja Sofrenovic , Kimberly McEwan , Suzanne Crowe , Jenelle Marier , Robbie Davies , Erik J. Suuronen1,2*, Drew Kuraitis 1,2* 1 Division of Cardiac Surgery, University of Ottawa Heart Institute, Ottawa, Canada, 2 Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Canada, 3 Department of Mechanical Engineering, University of Ottawa, Ottawa, Canada, 4 Department of Statistics, University of Ottawa Heart Institute, Ottawa, Canada Abstract Background: Cell transplantation for regenerative medicine has become an appealing therapeutic method; however, stem and progenitor cells are not always freshly available. Cryopreservation offers a way to freeze cells as they are generated, for storage and transport until required for therapy. This study was performed to assess the feasibility of cryopreserving peripheral blood mononuclear cells (PBMCs) for the subsequent in vitro generation of their derived therapeutic population, circulating angiogenic cells (CACs). Methods: PBMCs were isolated from healthy human donors. Freshly isolated cells were either analyzed immediately or cryopreserved in media containing 6% plasma serum and 5% dimethyl sulfoxide. PBMCs were thawed after being frozen for 1 (early thaw) or 28 (late thaw) days and analyzed, or cultured for 4 days to generate CACs. Analysis of the cells consisted of flow cytometry for viability and phenotype, as well as functional assays for their adhesion and migration potential, cytokine secretion, and in vivo angiogenic potential. Results: The viability of PBMCs and CACs as w