Establishment and Characterization of Four Long-Term Cultures of Neural Stem/Progenitor Cell Lines from the Japanese Flo
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Establishment and Characterization of Four Long-Term Cultures of Neural Stem/Progenitor Cell Lines from the Japanese Flounder Paralichthys olivaceus GAO Chen1), #, SONG Huayu1), 3), #, WANG Mengxun1), LIU Xiumei1), ZHAO Jun1), WANG Xubo1), and ZHANG Quanqi1), 2), * 1) Key Laboratory of Marine Genetics and Breeding, Ministry of Education, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China 2) Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China 3) Qingdao Conson Oceantec Valley Development Co., Ltd., Qingdao 266003, China (Received January 3, 2020; revised March 26, 2020; accepted May 8, 2020) © Ocean University of China, Science Press and Springer-Verlag GmbH Germany 2020 Abstract Neurogenesis is an important progress wherein the neural stem cells (NSCs) differentiate into functional neurons under conductive conditions. Neurogenesis occurs continuously in different areas of the central neural system in adult teleosts compared with adult mammals. Therefore, NSC cell lines must be established to offer a valuable in vitro system for studies on neurogenesis and other related functions. In this study, four cell lines designated as PoB1, PoB2, PoBf and PoBh were established from the brain of the Japanese flounder Paralichthys olivaceus. The cell lines were sub-cultured over 150 times and still grew well in DMEM/F12 medium at 24℃. PoB1, PoB2, PoBf and PoBh were identified as neural stem/progenitor cell lines on the basis of the mRNA expression of nestin and/or aldh1l1 or slc6a4 and the formation of neurospheres. The cells transfected with the pEGFP-C1 plasmid showed fluorescent signals with distinct reagent dependencies. The established cell lines from the brain of P. olivaceus offer a valuable system in vitro for the study of neurogenesis, fish neural regulation and endocrinology-related functions. Key words brain; cell lines; Paralichthys olivaceus; transfection; nestin
1 Introduction Neurogenesis, which mainly involves cell proliferation, migration, differentiation and survival, is an important progress that the neural stem cells (NSCs) differentiate into functional neurons under conductive conditions. Similar to other stem cells, NSCs also display self-renewal and differentiation abilities, as well as participate in the formation of the nervous system during development. In mammals, neural cells are mainly generated during the pre-natal phase of development, and neurogenesis is restricted to two regions of the adult brain (Taupin, 2006). However, neurogenesis considerably differs among various vertebrate classes. For example, teleosts continuously produce a tremendous number of new neurons in many regions of the adult brain. Moreover, the cell proliferation rate in teleosts is one to two orders of magnitude higher than that in mammals (Hinsch and Zupanc, 2006). Therefore, studying the NSCs in teleosts is important to understand the development and #
The two authors contributed equally
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