A Three-Dimensional Culture System with Matrigel Promotes Purified Spiral Ganglion Neuron Survival and Function In Vitro

Yan, Wenqing1,2; Liu, Wenwen1,2; Qi, Jieyu3,4,5; Fang, Qiaojun3,4,5; Fan, Zhaomin1,2; Sun, Gaoying1,2; Han, Yuechen1,2; Zhang, Daogong1,2; Xu, Lei1,2; Wang, Mingming1,2; Li, Jianfeng1,2; Chen, Fangyi6; Liu, Dong4; Chai, Renjie3,4,5; Wang, Haibo1,2

2018-03

10.1007/s12035-017-0471-0

MOLECULAR NEUROBIOLOGY   影响因子和分区

0893-7648

1559-1182

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.

Spiral ganglion neurons Three-dimensional culture Mutant mice Patch clamp

SCI

英语

其他

Natural Science Foundation of Jiangsu Province[BK20150022] ; Natural Science Foundation of Jiangsu Province[BK20140620] ; Natural Science Foundation of Jiangsu Province[BK20150598]

Neurosciences & Neurology

Neurosciences

WOS:000426897800021

HUMANA PRESS INC

NEUROSCIENCE & BEHAVIOR

Web of Science

1.Shandong Univ, Shandong Prov Hosp, Otolaryngol Head & Neck Surg, Jinan, Shandong, Peoples R China
2.Shandong Prov Key Lab Otol, Jinan, Shandong, Peoples R China
3.Southeast Univ, Inst Life Sci, Minist Educ, Key Lab Dev Genes & Human Dis, Nanjing 210096, Jiangsu, Peoples R China
4.Nantong Univ, Jiangsu Key Lab Neuroregenerat, Coinnovat Ctr Neuroregenerat, Qixiu Rd 19, Nantong 226001, Peoples R China
5.Southeast Univ, Zhongda Hosp, Dept Otolaryngol Head & Neck Surg, Nanjing 210096, Jiangsu, Peoples R China
6.South Univ Sci & Technol China, Dept Biol, Shenzhen, Peoples R China

Yan, Wenqing,Liu, Wenwen,Qi, Jieyu,et al. A Three-Dimensional Culture System with Matrigel Promotes Purified Spiral Ganglion Neuron Survival and Function In Vitro[J]. MOLECULAR NEUROBIOLOGY,2018,55(3):2070-2084.

Yan, Wenqing.,Liu, Wenwen.,Qi, Jieyu.,Fang, Qiaojun.,Fan, Zhaomin.,...&Wang, Haibo.(2018).A Three-Dimensional Culture System with Matrigel Promotes Purified Spiral Ganglion Neuron Survival and Function In Vitro.MOLECULAR NEUROBIOLOGY,55(3),2070-2084.

Yan, Wenqing,et al."A Three-Dimensional Culture System with Matrigel Promotes Purified Spiral Ganglion Neuron Survival and Function In Vitro".MOLECULAR NEUROBIOLOGY 55.3(2018):2070-2084.