title: A Three-Dimensional Culture System with Matrigel Promotes Purified Spiral Ganglion Neuron Survival and Function In Vitro.
abstract: In vitro culture of spiral ganglion neurons (SGNs) is a useful approach to investigate numerous aspects of neuronal behavior and to identify potential therapeutic targets for SGN protection and regeneration. However, the isolation of SGNs and the long-term maintenance of their structure and function in vitro remain challenging. In this study, we isolated SGNs from Bhlhb5-cre and Rosa26-tdTomato mice with fluorescence-activated cell sorting and determined the cell purity. We then encapsulated the pure SGNs in matrigel and cultured the SGNs in vitro. We found that the three-dimensional (3D)-matrigel culture environment significantly suppressed apoptosis and improved SGN survival in vitro, which enabled the long-term culture of SGNs for up to 6 months. The 3D-matrigel system also significantly promoted neurite outgrowth of the SGNs, increased the cells' polarity, promoted the area of growth cones, and significantly increased the synapse density of the SGNs. More importantly, the 3D-matrigel system helped to maintain and promote the electrophysiological properties of the SGNs. In conclusion, the 3D-matrigel culture system promoted the survival of purified SGNs in vitro and maintained their morphological structure and function and thus could be a useful tool for studying the physiology and pathophysiology of purified SGNs in long-term culture.
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Wenqing
authors.initials: W
authors.lastname: Yan
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Liu
authors.affiliation: Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Southeast University, Nanjing, 210096, China.|Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Qixiu Road 19, Nantong, 226001, China.|Department of Otolaryngology Head and Neck Surgery, Zhongda Hospital, Southeast University, Nanjing, 210096, China.
authors.forename: Jieyu
authors.initials: J
authors.lastname: Qi
authors.affiliation: Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Southeast University, Nanjing, 210096, China.|Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Qixiu Road 19, Nantong, 226001, China.|Department of Otolaryngology Head and Neck Surgery, Zhongda Hospital, Southeast University, Nanjing, 210096, China.
authors.forename: Qiaojun
authors.initials: Q
authors.lastname: Fang
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Zhaomin
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Gaoying
authors.initials: G
authors.lastname: Sun
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Yuechen
authors.initials: Y
authors.lastname: Han
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Daogong
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Mingming
authors.initials: M
authors.lastname: Wang
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.|Shandong Provincial Key Laboratory of Otology, Jinan, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Biology, South University of Science and Technology of China, Shenzhen, China.
authors.forename: Fangyi
authors.initials: F
authors.lastname: Chen
authors.affiliation: Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Qixiu Road 19, Nantong, 226001, China. liudongtom@gmail.com.
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.affiliation: Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Southeast University, Nanjing, 210096, China. renjiec@seu.edu.cn.|Co-innovation Center of Neuroregeneration, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Qixiu Road 19, Nantong, 226001, China. renjiec@seu.edu.cn.|Department of Otolaryngology Head and Neck Surgery, Zhongda Hospital, Southeast University, Nanjing, 210096, China. renjiec@seu.edu.cn.
authors.forename: Renjie
authors.initials: R
authors.lastname: Chai
authors.affiliation: Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China. wang.hb7585@hotmail.com.|Shandong Provincial Key Laboratory of Otology, Jinan, China. wang.hb7585@hotmail.com.
authors.forename: Haibo
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000008:administration & dosage*; D004338:Drug Combinations; D007797:Laminin / Q000008:administration & dosage*; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D011509:Proteoglycans / Q000008:administration & dosage*; D013136:Spiral Ganglion / Q000187:drug effects / Q000502:physiology*
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen
keywords: Mutant mice; Patch clamp; Spiral ganglion neurons; Three-dimensional culture