The Three-Dimensional Culture System with Matrigel and Neurotrophic Factors Preserves the Structure and Function of Spiral Ganglion NeuronIn Vitro

Author:

Sun Gaoying12,Liu Wenwen12,Fan Zhaomin1,Zhang Daogong1,Han Yuechen1,Xu Lei1,Qi Jieyu34,Zhang Shasha34,Gao Bradley T.5,Bai Xiaohui12,Li Jianfeng12,Chai Renjie34,Wang Haibo12

Affiliation:

1. Otolaryngology-Head and Neck Surgery, Provincial Hospital Affiliated to Shandong University, Jinan 250022, China

2. Shandong Provincial Key Laboratory of Otology, Jinan 250022, China

3. Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Southeast University, Nanjing 210096, China

4. Co-Innovation Center of Neuroregeneration, Nantong University, Nantong 226001, China

5. College of Medicine, The University of Tennessee Health Science Center, Memphis, TN 38163, USA

Abstract

Whole organ culture of the spiral ganglion region is a resourceful model system facilitating manipulation and analysis of live sprial ganglion neurons (SGNs). Three-dimensional (3D) cultures have been demonstrated to have many biomedical applications, but the effect of 3D culture in maintaining the SGNs structure and function in explant culture remains uninvestigated. In this study, we used the matrigel to encapsulate the spiral ganglion region isolated from neonatal mice. First, we optimized the matrigel concentration for the 3D culture system and found the 3D culture system protected the SGNs against apoptosis, preserved the structure of spiral ganglion region, and promoted the sprouting and outgrowth of SGNs neurites. Next, we found the 3D culture system promoted growth cone growth as evidenced by a higher average number and a longer average length of filopodia and a larger growth cone area. 3D culture system also significantly elevated the synapse density of SGNs. Last, we found that the 3D culture system combined with neurotrophic factors had accumulated effects in promoting the neurites outgrowth compared with 3D culture or NFs treatment only groups. Together, we conclude that the 3D culture system preserves the structure and function of SGN in explant culture.

Funder

National Basic Research Program of China

Publisher

Hindawi Limited

Subject

Neurology (clinical),Neurology

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