Damage to cochlear spiral ganglion neurons (SGNs) causes irreversible sensorineural hearing loss (SNHL), yet the underlying degenerative mechanisms remain elusive, impeding targeted therapies. Here, we established an auditory neuropathy model using the ototoxic drug ouabain to induce selective SGN injury. Transcriptomic profiling revealed a ferroptosis-linked gene signature, with functional assays confirming ferroptotic damage in SGNs. Pharmacological inhibition of ferroptosis mitigated SGN loss and hearing impairment. Lipidomics demonstrated dysregulated fatty acid metabolism with an elevated saturated-to-monounsaturated fatty acid ratio, accompanied by downregulation of stearoyl-CoA desaturase 1 (SCD1), the rate-limiting enzyme for monounsaturated fatty acid synthesis. Restoring SCD1 activity through SCD1 overexpression or monounsaturated fatty acid supplementation mitigated ouabain-induced ferroptosis in SGN and preserved auditory function, whereas genetic ablation of SCD1 aggravated degeneration. Bioinformatic screening and chromatin immunoprecipitation-PCR identified microphthalmia-associated transcription factor (MITF) as a direct upstream transcriptional regulator of SCD1. MITF overexpression restored SCD1 expression, suppressed ferroptosis, and improved SGN survival, while disruption of the MITF-SCD1 axis abrogated these protective effects, which were further validated in a cisplatin ototoxicity model. Collectively, these findings reveal a MITF-SCD1 lipid metabolic axis that safeguards SGNs against ferroptosis by sustaining SCD1-dependent lipid desaturation, highlighting a promising therapeutic target for SNHL.

Otitis media prevention remains challenging due to antibiotic resistance and frequent recurrence. This review proposes a novel strategy based on probiotic modulation of the Eustachian tube microenvironment. The approach leverages a dual-mechanism whereby probiotics secrete biosurfactants that contribute to the restoration of physiological surface tension and mucus flow, while simultaneously producing antimicrobial peptides that directly eliminate pathogens and suppress inflammatory responses. This synergistic action may not only facilitates sustainable colonization by beneficial bacteria but also disrupts the cycle of biofilm formation and functional impairment in the Eustachian tube. Experimental evidence and clinical observations indicate that this probiotic-based intervention correlates with significantly reduced incidence of otitis media and decreased recurrence rates. Future translation into clinical practice will require further development of optimized strain-specific formulations and effective delivery systems to fully realize this therapeutic approach. This review aims to critically evaluate current evidence supporting probiotic-mediated modulation of the Eustachian tube microenvironment, with particular emphasis on the complementary roles of biosurfactants and antimicrobial peptides in preventing otitis media, while highlighting the current limitations and future translational challenges. These findings highlight the potential of probiotic-mediated modulation of the Eustachian tube environment as a promising preventive strategy.

N6-methyladenosine (m6A) modification is a major post-transcriptional regulatory mechanism implicated in diverse biological and disease processes. However, its role in controlling ovarian follicle activation and female reproductive disorders remains poorly understood. WTAP, a component of the m6A methyltransferase complex, regulates the nuclear speckle localization of the stable heterodimer core complex. However, the specific role of WTAP in ovarian follicular development remains unclear. This study aimed to investigate the physiological functions and underlying mechanisms by which WTAP regulates oocyte maturation and follicular development. We generated oocyte-specific Wtap knockout mice and performed histological analysis, immunostaining, RNA-seq, and m6A-modified RNA immunoprecipitation sequencing (MeRIP-seq) on germinal vesicle (GV) and growing oocytes (GO). Reverse transcription quantitative real-time PCR (RT-qPCR) and immunostaining assessed primordial follicle activation; MeRIP-qPCR measured m6A enrichment on insulin receptor transcripts. Functional rescue experiments employed pharmacological inhibitors targeting the PI3K-AKT signaling pathway. The findings revealed that oocyte-specific deletion of WTAP impedes oocyte growth and causes female infertility. Loss of WTAP in oocytes triggers the premature activation of primordial follicles. Temporal transcriptome profiling revealed significant maternal RNA accumulation in Wtap knockout oocytes during growth. Analysis of m6A dynamics in GO indicated that WTAP-mediated m6A methylation ensures timely activation of the PI3K-AKT signaling pathway, which regulates the activation of primordial follicles. Moreover, we found that insulin receptor (INSR) is the key target of WTAP-dependent m6A modification. Crucially, pharmacological inhibition of INSR and phosphatidyqinositol‐3 kinase (PI3K) efficiently rescued follicular developmental defects caused by oocyte-specific WTAP deficiency. These findings identify a WTAP-m6A-INSR regulatory axis that links epitranscriptomic regulation to PI3K-AKT signaling during ovarian follicle development and suggest potential biomedical relevance for disorders characterized by abnormal follicle activation and female infertility.

Myostatin (MSTN), a member of the TGF-β superfamily, is recognized for its role in regulating muscle growth and cancer cachexia, but its involvement in glioma remains unclear. We examined MSTN expression and its clinical significance in glioma using public datasets and single-cell sequencing data. Immunohistochemistry and western blot further supported elevated MSTN protein levels in GBM tissues and cell lines. Knocking down MSTN in U87 and T98G cells revealed its impact on proliferation, migration, invasion, ROS accumulation, and apoptosis. Our analysis demonstrated that MSTN was significantly upregulated in glioma tissues and cell lines. Furthermore, ROC analysis indicated the diagnostic potential of MSTN in glioblastoma (GBM), lower-grade glioma (LGG), and the combined GBM-LGG cohort. High MSTN expression was associated with poorer survival in specific glioma subgroups, tumor heterogeneity, genomic instability-related features, and immune-related bioinformatic signatures. In vitro, siRNA-mediated MSTN depletion was associated with reduced glioblastoma cell proliferation, colony formation, migration, and invasion, together with increased ROS accumulation and apoptosis. Our findings suggest that MSTN upregulation in glioma could serve as a potential biomarker and might contribute to malignant phenotypes of glioma cells. Further studies are required to clarify the mechanistic role and clinical relevance of MSTN in glioma.

Background Genetic diagnosis plays a critical role in enhancing the identification and clinical management of pediatric hearing loss, ultimately improving both physiological outcomes and social-emotional development. As one of the most prevalent sensory disorders characterized by substantial genetic heterogeneity, hearing loss requires the implementation of efficient diagnostic approaches to inform targeted clinical interventions and personalized care strategies. Methods Here, we recruited 3353 Han Chinese children with nonsyndromic hearing loss, including 598 multiplex cases with family history. We employed SNPscan, targeted panel sequencing (TPS), and whole-exome sequencing (WES) to delineate the mutation landscape. Findings Utilizing a tiered genomic testing strategy, we performed SNPscan for all 3353 participants, TPS for 675 cases, and WES for 204 multiplex cases. We achieved an overall diagnostic yield of 48.3% (1620/3353) in this cohort. Our analysis identified 87 previously unreported variants across 24 genes, including a de novo FOXI1 mutation (c.479_481del) linked to Mondini malformation and enlarged vestibular aqueduct, validated through knock-in mouse models. Additionally, we discovered NEU4 as a candidate gene related to hearing loss, with knockout mice and cellular models demonstrating its role in auditory dysfunction via disrupted neuraminidase activity and axonal development. These findings enrich the spectrum of deafness-related genes and provide a new theoretical foundation for diagnosing hearing loss. Interpretation This study not only expands the genetic spectrum of hearing loss but also underscores the clinical utility of advanced sequencing in guiding precision therapies and early syndromic surveillance. This study offers a detailed characterization of the genetic landscape of pathogenic variants associated with hearing loss in the Han Chinese population, which will contribute to the design of a cost-effective genetic screening scheme, and also provides valuable insights that contribute to the global understanding of the genetics of hearing loss. Funding This work was supported by the Key Program of National Natural Science Foundation of China, the National Key Research and Development Program of China, the National Natural Science Foundation of China and the Major Fundamental Research Program of the Natural Science Foundation of Shandong Province, China.

Lung adenocarcinoma (LUAD) remains a leading cause of cancer-related death, underscoring the need for an improved molecular understanding. This study investigated the regulatory mechanism of the long non-coding RNA deoxyguanosine kinase antisense RNA 1 (DGUOK-AS1) in LUAD. DGUOK-AS1 was significantly upregulated in LUAD cells and serum samples, and its elevated expression showed a preliminary association with LUAD. Functional experiments demonstrated that DGUOK-AS1 promoted LUAD proliferation and migration both in vitro and in vivo, partly by acting as a competing endogenous RNA for miR-2467-5p to modulate PRMT5 expression. Mechanistically, RNA-binding motif protein 15 (RBM15) enhanced DGUOK-AS1 stability through m6A modification, which in turn enabled heterogeneous nuclear ribonucleoprotein H1 (HNRNPH1) binding in an m6A-dependent manner via its RNA recognition motif 3 (RRM3) domain, promoting degradation. RBM15 knockdown attenuated the malignant phenotype through the miR-2467-5p/PRMT5 axis. These findings reveal an m6A-dependent mechanism governing DGUOK-AS1 stability and provide insights into its contribution to LUAD progression.

Auditory neuropathy (AN) is a hearing disorder that is often overlooked due to its hidden cochlear activity and hearing loss profiles. Diphtheria toxin (DT) is commonly used to induce deafness in conditional gene knockout mouse models, but its ototoxic targets in wild-type animals remain controversial. This study aimed to characterize the pathogenic effects of DT on the cochlea of wild-type mice and establish a reliable model of AN. Adult CBA/CaJ mice of both sexes (n = 81) were administered by DT for three consecutive days. AN was confirmed by electrophysiological profiles: abnormal auditory brainstem response (ABR) with preserved distortion product otoacoustic emissions (DPOAE), followed by histopathological evaluation of the cochlear auditory pathway. DT induced a characteristic AN-characteristic hearing loss in mice. It initiated with ribbon synapse degeneration in inner hair cells (IHCs) within 24 h, coinciding with a significant reduction in ABR amplitude peak I. By post-injection day 3, extensive damage was observed, including IHC loss, degeneration of type I spiral ganglion neurons (SGNs), and demyelination of their axons. Despite a minimal loss of OHCs ( 2.1

Intracellular calcium increase and neurotransmitter release in vestibular hair cells (VHCs) play central roles in equilibrioception, which is one of the basic senses essential for daily life activities and movement in mammals. Independent of mechano-electrical transduction (MET), whether Gq protein-coupled receptor (GqPCR) signaling participate in the regulation of intracellular calcium dynamics and induce neuronal transmitter release in hair cells remains unknown. We screened mechanosensitive GqPCRs in VHCs and found that a Class C GPCR, metabotropic glutamate receptor 2 (mGlu2), is expressed in kinocilia and is essential for normal balance. Notably, the dispensable role of mGlu2 in normal hearing is consistent with absent of mature kinocilia in cochlea hair cells. Different from the conventional mGlu2-Gi signaling, the sensing of mechanical signals by mGlu2 activates the Gq‒PLCD4 pathway, increases intracellular calcium concentration and promotes neurotransmitter release in VHCs. Hair cell-specific deficiency of either Grm2 or Gnaq , or knockdown of Plcd4 expression, but not deficiency of another GqPCR Gpr68 , causes significant balance deficits. Reintroduction of mGlu2 into the VHCs of Pou4f3-CreER +/− Grm2 fl/fl mice restore vestibular functions. Our study reveals a previously uncharacterized role of GPCR signaling in equilibrioception and provides important insight into kinocilia signaling in VHCs, which are absent in mature cochlear hair cells.

Degeneration of cochlear spiral ganglion neurons (SGNs) leads to irreversible sensorineural hearing loss (SNHL), as SGNs lack regenerative capacity. Although cochlear glial cells (GCs) have some neuronal differentiation potential, their specific identities remain unclear. This study identifies a distinct subpopulation, Frizzled10 positive (FZD10+) cells, as an important type of GC responsible for neuronal differentiation in mouse cochlea. FZD10 + cells can differentiate into various SGN subtypes in vivo, adhering to natural proportions. Wnt signaling enhances the ability of FZD10 + cells to function as neural progenitors and increases the neuronal excitability of the FZD10-derived neurons. Single-cell RNA sequencing analysis characterizes FZD10-derived differentiating cell populations, while crosstalk network analysis identifies multiple signaling pathways and target genes influenced by Wnt signaling that contribute to the function of FZD10 + cells as neural progenitors. Pseudotime analysis maps the differentiation trajectory from proliferated GCs to differentiating neurons. Further experiments indicate that glypican 6 (GPC6) may regulate this neuronal lineage, while GPC6 deficiency diminishes the effects of Wnt signaling on FZD10-derived neuronal differentiation and synapse formation. These findings suggest the critical role of Wnt signaling in the neuronal differentiation derived from cochlear FZD10 + cells and provide insights into the mechanisms potentially involved in this process.

Human genome analyses have revealed that abnormal BAF (BRG1/BRM-associated factor) complex is highly associated with hearing loss. However, the underlying pathogenesis remains largely unknown. Disrupted structure and function of the organ of Corti is the most prevalent cause of sensorineural hearing loss in mammals. Here, we investigated the role of Brg1-based BAF complex during the differentiation and development of the auditory sensory epithelium, a crucial period for the formation of the organ of Corti. Our findings indicate that deletion of Brg1 leads to premature hair cell (HC) differentiation by inactivating Sonic hedgehog (Shh) signaling. Despite the formation of HCs, subsequent differentiation of inner hair cells (IHCs) and outer hair cells (OHCs) was impaired. Additionally, we observed that the mosaic-like arrangement of HCs and supporting cells (SCs) was disrupted resulting in abnormal sensory epithelium patterning. Furthermore, we found the planar cell polarity of the Brg1-deficient cochlea was abnormal. Our study demonstrates the pivotal role of Brg1 in the differentiation and patterning of the organ of Corti.

Severe to profound sensorineural hearing loss seriously affects the communication and cognitive ability of the patients. Cochlear implantation (CI) is currently the most effective treatment, while it may damage the remaining inner ear function due to its poor biocompatibility and the resultant fibrosis. Herein, a porous methacrylated poly(dimethylsiloxane) (MA-PDMS)-coated cochlear electrode is presented for CI and hearing protection. The porous MA-PDMS is filled with a hybrid hydrogel system made of dexamethasone sodium phosphate (Dex), Ti3C2Tx MXene (MXene), and methacrylate gelatin (GelMA). The coating shows good biocompatibility and drug loading and release capacity in vitro, protective effects on hair cells (HCs) and spiral ganglion neurons (SGNs) of the inner ear, as well as the residual hearing protection and the effective fibrosis reduction in vivo. It is anticipated that this porous electrode drug-loading coating may provide a valuable reference strategy for the future cochlear electrode transplantation system.

Spiral ganglion neurons (SGNs) in the inner ear are indispensable for auditory function, and their irreversible damage causes permanent sensorineural hearing loss. Although current human pluripotent stem cell (hPSC)-derived otic lineages offer a valuable resource for SGN regeneration, they face challenges in terms of reproducibility and functional maturation. Here, a robust protocol is established to generate human otic neuronal spheroids (hONS) from cryopreserved hPSC-derived pre-placodal ectoderm (PPE) cells. Post-thaw PPE cells retained high purity and differentiation efficiency comparable to fresh PPE cells. These self-assembled hONS differentiated into functionally mature SGN-like neurons, showing specific maker expression, electrophysiological activity, AMPA receptor-mediated glutamate response, and extensive neurite extension. In tripartite cocultures incorporating murine cochlear explants and human cortical organoids, hONS formed bidirectional functional synaptic connections, validated through live-cell imaging, optogenetic stimulation, and synaptic immunostaining. Notably, hONS exhibited heightened sensitivity to ototoxic insults. Short-term cisplatin exposure induced dose-dependent alterations in cellular and calcium dynamics, whereas prolonged exposure impaired glutamatergic neural functionality and triggered progressive neuronal death. Co-treatment with sodium thiosulfate attenuated cisplatin-induced damage. The hONS model also demonstrated concentration-dependent toxicity to neomycin. Collectively, this hONS model provides a reliable platform for investigating SGN regeneration and conducting preclinical evaluation of ototoxic drug.

Equilibrioception (sensing of balance) is essential for mammals to perceive and navigate the three-dimensional world. A rapid mechanoelectrical transduction (MET) response in vestibular hair cells is crucial for detecting position and motion. Here, we identify the G protein-coupled receptor (GPCR) LPHN2/ADGRL2, expressed on the apical membrane of utricular hair cells, as essential for maintaining normal balance. Loss of LPHN2 specifically in hair cells impaired both balance behavior and the MET response in mice. Functional analyses using hair-cell-specific Lphn2-knockout mice and an LPHN2-specific inhibitor suggest that LPHN2 regulates tip-link-independent MET currents at the apical surface of utricular hair cells. Mechanistic studies in a heterologous system show that LPHN2 converts force stimuli into increased open probability of transmembrane channel-like protein 1 (TMC1). LPHN2-mediated force sensation triggers glutamate release and calcium signaling in utricular hair cells. Importantly, reintroducing LPHN2 into the hair cells of Lphn2-deficient mice restores vestibular function and MET response. Our data reveal that a mechanosensitive GPCR is required for equilibrioception.

Hearing is one of the most vital sensory functions in human beings and a crucial means of perceiving and acquiring information from the natural environment. The advancement of human society is closely linked to the development of language, with hearing serving as the foundation for verbal communication. As individuals age, the deterioration of the auditory system becomes a significant factor contributing to sensory impairments in the elderly. In addition to hearing loss, the aging of the auditory system is also associated with cognitive decline and psychosocial disorders, which severely impact the quality of life for older adults. Currently, there are no effective treatments or interventions available for addressing the aging of the auditory system. Therefore, identifying biomarkers of the auditory system aging is of great significance. The Aging Biomarker Consortium of China has conducted a comprehensive evaluation of aging biomarkers in the auditory system, focusing on three dimensions: morphological, functional, and humoral biomarkers. This initiative aims to establish a foundation for assessing the degree of aging in the auditory system and to promote the management of auditory health in an aging society, ultimately enhancing the auditory health of the elderly population both in China and globally.

Mammalian spiral ganglion neurons (SGNs) in the cochlear are crucial for auditory signal processing. The degeneration and loss of SGNs leads to irreversible sensorineural hearing loss (SNHL) due to their limited regenerative capacity. However, the anatomical complexity and restricted accessibility of SGNs pose challenges for their research. In this study, we established a conditionally immortalized cell line, Shandong Institute of Otorhinolaryngology-spiral ganglion neuron 1 (SIO-SGN1), by introducing SV40 large T antigen into neonatal mouse SGNs. SIO-SGN1 cells showed robust proliferative capability and maintained high viability over 20 passages. They exhibited contact inhibition and expressed neuronal-specific markers but not glial or hair cell markers. Transcriptome analysis revealed that the transcriptomic profile of SIO-SGN1 cells closely resembles that of primary SGNs at embryonic day 15.5 and postnatal day 1. These cells highly expressed genes related to neuron development, axon guidance, synapse formation, and stemness. Treatment with the ototoxic drugs cisplatin or ouabain caused significant cell loss and damage in SIO-SGN1 cells, which was consistent with the drug responses observed in cultured primary SGNs. Collectively, our findings suggest that SIO-SGN1 cells serve as a promising in vitro model for screening ototoxic and otoprotective drugs, and investigating the molecular mechanisms underlying ototoxic drug-induced SGN loss and hearing impairment.

The conversion of force sensation into electrical signals via mechano-electrical transduction (MET) is considered the key step in auditory perception. Here, we find that the G-protein-coupled receptor (GPCR) LPHN2/ADGRL2 is expressed at the tips of stereocilia in cochlear hair cells and is associated with MET channel components. Hair-cell-specific LPHN2 deficiency causes hearing loss and impaired MET responses. A specific inhibitor of LPHN2 also reversibly blocks the MET response. Mechanistically, the administration of force to LPHN2 activates TMC1 through physical interaction and causes conformational changes in TMC1. Furthermore, the sensing of force by LPHN2 stimulates the Ca2+ response and neurotransmitter release in hair cells. Finally, expression of LPHN2-GAIN in cochlear hair cells of Lphn2-deficient mice prevents hearing loss. Our work provides evidence that the GPCR can play modulatory roles in the auditory process through a TMC1-coupled mechanism.

Mammalian cochlea spiral ganglion neurons (SGNs) are crucial for sound transmission, they can be damaged by chemotherapy drug cisplatin and lead to irreversible sensorineural hearing loss (SNHL), while such damage can also render cochlear implants ineffective. However, the mechanisms underlying cisplatin-induced SGNs damage and subsequent SNHL are still under debate and there is no currently effective clinical treatment. Here, this study demonstrates that ferroptosis is triggered in SGNs following exposure to cisplatin. Inhibiting ferroptosis protects against cisplatin-induced SGNs damage and hearing loss, while inducing ferroptosis intensifies these effects. Furthermore, cisplatin prompts nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy in SGNs, while knocking down NCOA4 mitigates cisplatin-induced ferroptosis and hearing loss. Notably, the upstream regulator of NCOA4 is identified and transcription factor forkhead box O1 (FOXO1) is shown to directly suppress NCOA4 expression in SGNs. The knocking down of FOXO1 amplifies NCOA4-mediated ferritinophagy, increases ferroptosis and lipid peroxidation, while disrupting the interaction between FOXO1 and NCOA4 in NCOA4 knock out mice prevents the cisplatin-induced SGN ferroptosis and hearing loss. Collectively, this study highlights the critical role of the FOXO1-NCOA4 axis in regulating ferritinophagy and ferroptosis in cisplatin-induced SGNs damage, offering promising therapeutic targets for SNHL mitigation.

Hereditary deafness is a common neurosensory disorder, and 148 non-syndromic deafness genes have been identified to date. Gene therapy has been used to treat a variety of genetic diseases, but no gene therapy drug for hereditary deafness has been approved for clinical use. At present, several clinical trials of gene therapy for hereditary deafness are underway. However, few normative documents have been issued to guide the standardization of gene therapy for hearing loss, and this document is the first global gene therapy guideline for hereditary hearing loss. The guidelines were jointly developed and drafted by experienced audiologists, virologists and biologists who are vigorously involved in inner ear gene therapy research in the Hearing, Speech and Communication Subsociety of Biophysical Society of China, Audiology Development Foundation Of China and Audiology Subsociety of Jiangsu Medical Association. These guidelines cover preclinical research and clinical practice of gene therapy for hereditary deafness, including indications, key points of pre-clinical research, patient selection criteria, pre-clinical preparation, drug efficacy, drug safety evaluation criteria, ethical review, etc. We hope that the guidelines will promote the standardization of clinical practice related to gene therapy for hereditary deafness in China and around the world.

Inner ear cell regeneration from stem/progenitor cells provides potential therapeutic strategies for the restoration of sensorineural hearing loss (SNHL), however, the efficiency of regeneration is low and the functions of differentiated cells are not yet mature. Biomaterials have been used in inner ear cell regeneration to construct a more physiologically relevant 3D culture system which mimics the stem cell microenvironment and facilitates cellular interactions. Currently, these biomaterials include hydrogel, conductive materials, magneto-responsive materials, photo-responsive materials, etc. We analyzed the characteristics and described the advantages and limitations of these materials. Furthermore, we reviewed the mechanisms by which biomaterials with different physicochemical properties act on the inner ear cell regeneration and depicted the current status of the material selection based on their characteristics to achieve the reconstruction of the auditory circuits. The application of biomaterials in inner ear cell regeneration offers promising opportunities for the reconstruction of the auditory circuits and the restoration of hearing, yet biomaterials should be strategically explored and combined according to the obstacles to be solved in the inner ear cell regeneration research.

Spiral ganglion neurons (SGNs) in the mammalian cochleae are essential for the delivery of acoustic information, and damage to SGNs can lead to permanent sensorineural hearing loss as SGNs are not capable of regeneration. Cochlear glial cells (GCs) might be a potential source for SGN regeneration, but the neuronal differentiation ability of GCs is limited and its properties are not clear yet. Here, we characterized the cochlear Sox10-positive (Sox10+) GCs as a neural progenitor population and developed a basement membrane extract-based three-dimensional (BME-3D) culture system to promote its neuronal generation capacity in vitro. Firstly, the purified Sox10+ GCs, isolated from Sox10-creER/tdTomato mice via flow cytometry, were able to form neurospheres after being cultured in the traditional suspension culture system, while significantly more neurospheres were found and the expression of stem cell-related genes was upregulated in the BME-3D culture group. Next, the BME-3D culture system promoted the neuronal differentiation ability of Sox10+ GCs, as evidenced by the increased number, neurite outgrowth, area of growth cones, and synapse density as well as the promoted excitability of newly induced neurons. Notably, the BME-3D culture system also intensified the reinnervation of newly generated neurons with HCs and protected the neurospheres and derived-neurons against cisplatin-induced damage. Finally, transcriptome sequencing analysis was performed to identify the characteristics of the differentiated neurons. These findings suggest that the BME-3D culture system considerably promotes the proliferation capacity and neuronal differentiation efficiency of Sox10+ GCs in vitro, thus providing a possible strategy for the SGN regeneration study.