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Cathepsin L involved in the freezing resistance of murine normal hatching embryos and dormant embryos
Affiliation:1. College of Animal Science and Technology, Beijing University of Agriculture, Beijing, 102206, China;2. College of Economics and Management, Beijing University of Agriculture, Beijing, 102206, China;3. China National Biotec Group Company Limited, Beijing, 102206, China;1. Department of Laboratory, The Affiliated Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu, China;2. Department of Gynecology, The Affiliated Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu, China;1. Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, Istanbul, Turkey;2. Cellular and Molecular Therapeutics Branch, National Heart, Lung, and Blood Institute (NHLBI)/National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD, USA;1. Ojinmed IVF Center, Ulaanbaatar, Mongolia;2. Department of Molecular Biology and Genetics, School of Biomedicine, Mongolian National University of Medical Sciences, Ulaanbaatar, Mongolia;3. Department of Biochemistry, School of Medicine, International University of Health and Welfare, Narita, Japan;1. Laboratory of Genetics and Evolution, Institute of Zoology and Biomedical Research, Jagiellonian University, Krakow, Poland;2. Forensic Genetics Section, Institute of Forensic Research, Krakow, Poland;3. Deptartment of Molecular Neuropharmacology, Institute of Pharmacology, Polish Academy of Sciences, Krakow, Poland;4. Max-Delbrück-Center for Molecular Medicine (MDC) in the Helmholtz Association, Berlin, Germany;5. Department of Molecular Biology, Institute of Genetics and Animal Biotechnology, Polish Academy of Sciences Jastrz?biec, Magdalenka, Poland;1. Department of Obstetrics and Gynecology, Second Xiangya Hospital, Central South University, Changsha, 410011, China;2. Department of Clinical Medicine, Harbin Medical University, Harbin, 150081, China;1. Department of Gynecologic Oncology, Shaanxi Provincial Cancer Hospital, Xi’an 710061, China;2. Department of Gastrointestinal Oncology, Shaanxi Provincial Cancer Hospital, Xi’an 710061, China
Abstract:The cryopreservation of mammalian embryos is an important technology in embryo engineering. The discovery and application of the embryo's own high freezing resistance factors are the main methods to improve the utilization of mammalian embryos in cryopreservation. Cathepsin L gene expression in the frozen and thawed dormant embryos displayed a significant difference from those normal hatched ones. The aim of the present study was to dig out the potential role of Cathepsin L in anti-freezing capacity of murine blastocysts by investigating the location and expression of Cathepsin L in frozen and thawed both activated and dormant hatching blastocysts. Different concentrations of Cathepsin L recombinant protein and E-64d were then respectively added into the embryo cryoprotectant and pre-cryo culture medium. Our results found that down-regulation of Cathepsin L improves the freezing resistance of murine normal hatching embryos by reducing apoptosis. Cathepsin L inhibitors can be used to improve the efficiency of cryopreservation and recovery of blastocysts in vitro. Our study provides a theoretical basis for the further development and application of Cathepsin L.
Keywords:Mouse  Dormant embryos  Hatching blastocyst  Cryopreservation  Cathepsin L
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