Pinacidil enhances survival of cryopreserved human embryonic stem cells |
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| Institution: | 1. Centre for Stem Cell Biology, University of Sheffield, Western Bank, Sheffield S10 2TN, UK;2. Sheffield Diagnostic Genetic Services, Sheffield Children’s Hospital, Sheffield S10 2TH, UK;1. Key Laboratory of Poyang Lake Ecology and Bio-Resource Utilization of Ministry of Education, Nanchang University, Nanchang 330031, China;2. Department of Chemical Engineering, Nanchang University, Nanchang 330031, China;3. School of Foreign Language, Nanchang University, Nanchang 330031, China;1. Department of Anatomy, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan;2. Department of Nutritional Science, Okayama Prefectural University, 111 Kuboki, Soja, Okayama, 719-1197, Japan;3. Department of Anatomy and Cell Biology, Institute of Health Biosciences, The University of Tokushima, Graduate School, 3-18-15 Kuramoto, Tokushima, 770-8503, Japan;1. Haartman Institute, Department of Bacteriology and Immunology, and Research Programs Unit, Immunobiology, University of Helsinki, PB 21, FIN-00014 University of Helsinki, Finland;2. Department of Surgery, Hospital for Children and Adolescents, Helsinki University Hospital, PB 281, FIN-00029 HUS, Finland;1. Departamento de Biología Funcional y Antropología Física, Universitat de València, Burjassot, 46100 Valencia, Spain;2. Centro de Tecnología Animal, Instituto Valenciano de Investigaciones Agrarias (CITA-IVIA), Apdo 187, Pol. La Esperanza n°100, 12400 Segorbe, Castellón, Spain;3. TECNOGAM Research Group, Instituto Universitario de Ciencias Ambientales (IUCA), Departamento de Producción Animal y Ciencia de los Alimentos, Universidad de Zaragoza, Huesca, Spain;1. Center for Research and Development of Agricultural Industry, Faculty of Agro Industry Product and Innovation Technology, Srinakarinwirot University, Ongkarak 26120, Thailand;2. Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Hiroshima 739-8530, Japan |
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| Abstract: | Human embryonic stem cells (hESCs) can be maintained as undifferentiated cells in vitro and induced to differentiate into a variety of somatic cell types. Thus, hESCs provide a source of differentiated cell types that could be used to replace diseased cells of a tissue. The efficient cryopreservation of hESCs is important for establishing effective stem cell banks, however, conventional slow freezing methods usually lead to low rates of recovery after thawing cells and their replating in culture. We have established a method for recovering cryopreserved hESCs using pinacidil and compared it to a method that employs the ROCK inhibitor Y-27632. We show that pinacidil is similar to Y-27632 in promoting survival of hESCs after cryopreservation. The cells exhibited normal hESC morphology, retained a normal karyotype, and expressed characteristic hESC markers (OCT4, SSEA3, SSEA4 and TRA-1-60). Moreover, the cells retained the capacity to differentiate into derivatives of all three embryonic germ layers as demonstrated by differentiation through embryoid body formation. Pinacidil has been used for many years as a vasodilator drug to treat hypertension and its manufacture and traceability are well defined. It is also considerably cheaper than Y-27632. Thus, the use of pinacidil offers an efficient method for recovery of cryopreserved dissociated human ES cells. |
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