title: Prevention of Osmotic Injury to Human Umbilical Vein Endothelial Cells for Biopreservation: A First Step Toward Biobanking of Endothelial Cells for Vascular Tissue Engineering.
abstract: High-survival-rate cryopreservation of endothelial cells plays a critical role in vascular tissue engineering, while optimization of osmotic injuries is the first step toward successful cryopreservation. We designed a low-cost, easy-to-use, microfluidics-based microperfusion chamber to investigate the osmotic responses of human umbilical vein endothelial cells (HUVECs) at different temperatures, and then optimized the protocols for using cryoprotective agents (CPAs) to minimize osmotic injuries and improve processes before freezing and after thawing. The fundamental cryobiological parameters were measured using the microperfusion chamber, and then, the optimized protocols using these parameters were confirmed by survival evaluation and cell proliferation experiments. It was revealed for the first time that HUVECs have an unusually small permeability coefficient for Me2SO. Even at the concentrations well established for slow freezing of cells (1.5 M), one-step removal of CPAs for HUVECs might result in inevitable osmotic injuries, indicating that multiple-step removal is essential. Further experiments revealed that multistep removal of 1.5 M Me2SO at 25°C was the best protocol investigated, in good agreement with theory. These results should prove invaluable for optimization of cryopreservation protocols of HUVECs.
authors.affiliation: 1 Department of Electronic Science and Technology, Center for Biomedical Engineering, University of Science and Technology of China , Hefei, Anhui, China .
authors.forename: Dan
authors.initials: D
authors.lastname: Niu
authors.affiliation: 1 Department of Electronic Science and Technology, Center for Biomedical Engineering, University of Science and Technology of China , Hefei, Anhui, China .|2 Anhui Provincial Engineering Technology Research Center for Biopreservation and Artificial Organs , Hefei, Anhui, China .
authors.forename: Gang
authors.initials: G
authors.lastname: Zhao
authors.affiliation: 1 Department of Electronic Science and Technology, Center for Biomedical Engineering, University of Science and Technology of China , Hefei, Anhui, China .
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Liu
authors.affiliation: 2 Anhui Provincial Engineering Technology Research Center for Biopreservation and Artificial Organs , Hefei, Anhui, China .|3 Center for Reproductive Medicine, The Affiliated Hospital of Anhui Medical University , Hefei, Anhui, China .
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
authors.affiliation: 2 Anhui Provincial Engineering Technology Research Center for Biopreservation and Artificial Organs , Hefei, Anhui, China .|3 Center for Reproductive Medicine, The Affiliated Hospital of Anhui Medical University , Hefei, Anhui, China .
authors.forename: Yunxia
authors.initials: Y
authors.lastname: Cao
mesh_terms: D018070:Biological Specimen Banks; D001808:Blood Vessels / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D003451:Cryoprotective Agents / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000473:pathology*; D006801:Humans; D009995:Osmosis / Q000187:drug effects; D010477:Perfusion; D010539:Permeability; D011309:Preservation, Biological; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D003451:Cryoprotective Agents