Characterization of Rat Spiral Ligament Cell Line Immortalized by Adenovirus 12-Simian Virus 40 Hybrid Virus

Yian, Christopher H.; Moon, Sangook; Jin, Sunji; Webster, Paul; Rhim, Johng S.; Andalibi, Ali; Lim, David J.

doi:10.1177/000348940611501213

Cited by 9 publications

(11 citation statements)

References 43 publications

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“…Inner ear cell culture has been successfully reported in epithelial (Rarey and Patterson,1989; Achouche et al,1991; Melichar and Gitter,1992) and endothelial (Bowman et al,1985) cells. Recently, cell culture of SLFs has also been established and characterized for type I SLFs in gerbils (Gratton et al,1996), mice (Suko et al,2000), and rats (Yian et al,2006). To date, however, there are no reports on the successful establishment of a concurrent culture of all types of SLFs, which more closely mimics the in vivo condition.…”

Section: Discussionmentioning

confidence: 99%

Functional interaction between mesenchymal stem cells and spiral ligament fibrocytes

Sun

Fujisawa

Matsunaga

2012

J of Neuroscience Research

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Spiral ligament fibrocytes (SLFs) play an important role in normal hearing as well as in several types of sensorineural hearing loss attributable to inner ear homeostasis disorders. Our previous study showed that transplantation of mesenchymal stem cells (MSCs) into the inner ear of rats with damaged SLFs significantly accelerates hearing recovery compared with rats without MSC transplantation. To elucidate this mechanism of SLF repair and to determine the contribution of transplanted MSCs in this model, we investigated the mutual effects on differentiation and proliferation between MSCs and SLFs in a coculture system. Factors secreted by SLFs had the ability to promote the transdifferentiation of MSCs into SLF-like cells, and the factors secreted by MSCs had a stimulatory effect on the proliferation of SLFs. Cytokine antibody array analysis revealed the involvement of transforming growth factor-β (TGF-β) in SLF proliferation induced by MSCs. In addition, a TGF-β inhibitor reduced SLF proliferation induced by MSC stimulation. Our results suggest that there are two mechanisms of hearing recovery following transplantation of MSCs into the inner ear: 1) MSCs transdifferentiate into SLF-like cells that compensate for lost SLFs, and 2) transplanted MSCs stimulate the regeneration of host SLFs. Both mechanisms contribute to the functional recovery of the damaged SLF network.

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Section: Discussionmentioning

confidence: 99%

Functional interaction between mesenchymal stem cells and spiral ligament fibrocytes

Sun

Fujisawa

Matsunaga

2012

J of Neuroscience Research

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“…The rat SLF cell line, immortalized with adenovirus type 12-simian virus 40 hybrid virus (96), was maintained in Dulbecco's modified Eagle's medium (DMEM) (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum, penicillin (100 units/ml), and streptomycin (0.1 mg/ml). Epidermal fibroblasts of Sprague-Dawley rat (CRL-1213) were purchased from the ATCC (Manassas, VA).…”

Section: Methodsmentioning

confidence: 99%

“…Also, SLF cell lines (96) showed an induction in monocyte chemotactic protein 1 (MCP-1) expression after treatment with lysate of nontypeable Haemophilus influenzae (NTHI), one of the most common OM pathogens (72). Moreover, it has previously been shown that monocytes can infiltrate cochlea exhibiting chronic middle ear inflammation or acoustic trauma (22,34,37).…”

Section: -Derived Slfs Revealed Involvement Of Tlr2 and Myd88 In Nthimentioning

confidence: 99%

Toll-Like Receptor 2-Dependent NF-κB Activation Is Involved in Nontypeable Haemophilus influenzae -Induced Monocyte Chemotactic Protein 1 Up-Regulation in the Spiral Ligament Fibrocytes of the Inner Ear

et al. 2007

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Inner ear dysfunction secondary to chronic otitis media (OM), including high-frequency sensorineural hearing loss or vertigo, is not uncommon. Although chronic middle ear inflammation is believed to cause inner ear dysfunction by entry of OM pathogen components or cytokines from the middle ear into the inner ear, the underlying mechanisms are not well understood. Previously, we demonstrated that the spiral ligament fibrocyte (SLF) cell line up-regulates monocyte chemotactic protein 1 (MCP-1) expression after treatment with nontypeable Haemophilus influenzae (NTHI), one of the most common OM pathogens. We hypothesized that the SLF-derived MCP-1 plays a role in inner ear inflammation secondary to OM that is responsible for hearing loss and dizziness. Antibiotics have led to a dramatic decline in the incidence of life-threatening complications of otitis media (OM), such as meningitis or brain abscess (3). However, inner ear dysfunction secondary to chronic OM, including high-frequency sensorineural hearing loss or vertigo, is not uncommon (13,26,36,55,60). Although chronic middle ear inflammation is believed to cause inner ear dysfunction by entry of OM pathogen components or cytokines from the middle ear into the inner ear, the underlying mechanisms are not well understood (18,32,39,44,52,87).The inner ear is a sensory organ for hearing (cochlea) and equilibrium (vestibule). It consists of a variety of specialized cell types (50, 51), such as sensory hair cells, supporting cells, sulcus cells, and spiral ligament fibrocytes (SLFs), which are the most abundant cell types exposed to the perilymph. The type of inner ear cells that respond to proinflammatory signals entering the inner ear remain unknown. Considering that SLFs are one of the abundant cell types in the cochlea and that they secrete cytokines and chemokines after proinflammatory stimuli (72, 97), we hypothesized that the SLFs are major responders to such signals.Preliminary studies of human temporal bones with labyrinthitis showed the infiltration of lysozyme-positive round cells with a monomorphic nucleus into the spiral ligament (unpublished data). Also, SLF cell lines (96) showed an induction in monocyte chemotactic protein 1 (MCP-1) expression after treatment with lysate of nontypeable Haemophilus influenzae (NTHI), one of the most common OM pathogens (72). Moreover, it has previously been shown that monocytes can infiltrate cochlea exhibiting chronic middle ear inflammation or acoustic trauma (22,34,37). These results led us to focus on MCP-1 as an SLF-derived proinflammatory chemokine attracting effector cells and causing inner ear dysfunction.MCP-1, also known as the chemokine C-C motif ligand 2, is produced by various cells, including endothelial cells, smooth muscle cells, fibroblasts, and macrophages, in response to cytokines, growth factors, or bacterial components (9,46,78). It is encoded by an immediate-early gene (33) and is up-regulated by various stimuli such as bacterial lipopolysaccharide (LPS), interleukin-1 (IL-1), tumor necrosis fact...

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“…Rat spiral ligament fibrocyte cell line (RSL) were maintained in DMEM supplemented with 10% FBS, penicillin (100 U/ml), and streptomycin (0.1 mg/ml; Life Technologies) at 37°C in a humidified atmosphere of 5% CO 2 and 95% air as described (Yian et al, 2006 ). HEI-OC1 cells were cultured under a permissive condition (33°C, 10% CO 2 ) in high-glucose DMEM containing 10% FBS without antibiotics as described (Kalinec et al, 2003 ).…”

Section: Methodsmentioning

confidence: 99%

Interleukin-10 Attenuates Hypochlorous Acid-Mediated Cytotoxicity to HEI-OC1 Cochlear Cells

Mwangi

Kil

Phak

et al. 2017

Front. Cell. Neurosci.

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Inflammatory reaction plays a crucial role in the pathophysiology of acquired hearing loss such as ototoxicity and labyrinthitis. In our earlier work, we showed the pivotal role of otic fibrocytes in cochlear inflammation and the critical involvement of proinflammatory cytokines in cisplatin ototoxicity. We also demonstrated that otic fibrocytes inhibit monocyte chemoattractant protein 1 (CCL2) upregulation in response to interleukin-10 (IL-10) via heme oxygenase 1 (HMOX1) signaling, resulting in suppression of cochlear inflammation. However, it is still unclear how IL-10 affects inflammation-mediated cochlear injury. Here we aim to determine how hypochlorous acid, a model inflammation mediator affects cochlear cell viability and how IL-10 affects hypochlorous acid-mediated cochlear cell injury. NaOCl, a sodium salt of hypochlorous acid (HOCl) was found to induce cytotoxicity of HEI-OC1 cells in a dose-dependent manner. Combination of hydrogen peroxide and myeloperoxidase augmented cisplatin cytotoxicity, and this synergism was inhibited by N-Acetyl-L-cysteine and ML-171. The rat spiral ligament cell line (RSL) appeared to upregulate the antioxidant response element (ARE) activities upon exposure to IL-10. RSL cells upregulated the expression of NRF2 (an ARE ligand) and NR0B2 in response to CoPP (a HMOX1 inducer), but not to ZnPP (a HMOX1 inhibitor). Adenovirus-mediated overexpression of NR0B2 was found to suppress CCL2 upregulation. IL-10-positive cells appeared in the mouse stria vascularis 1 day after intraperitoneal injection of lipopolysaccharide (LPS). Five days after injection, IL-10-positive cells were observed in the spiral ligament, spiral limbus, spiral ganglia, and suprastrial area, but not in the stria vascularis. IL-10R1 appeared to be expressed in the mouse organ of Corti as well as HEI-OC1 cells. HEI-OC1 cells upregulated Bcl-xL expression in response to IL-10, and IL-10 was shown to attenuate NaOCl-induced cytotoxicity. In addition, HEI-OC1 cells upregulated IL-22RA upon exposure to cisplatin, and NaOCl cytotoxicity was inhibited by IL-22. Taken together, our findings suggest that hypochlorous acid is involved in cochlear injury and that IL-10 potentially reduces cochlear injury through not only inhibition of inflammation but also enhancement of cochlear cell viability. Further studies are needed to determine immunological characteristics of intracochlear IL-10-positive cells and elucidate molecular mechanisms involved in the otoprotective activity of IL-10.

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Characterization of Rat Spiral Ligament Cell Line Immortalized by Adenovirus 12-Simian Virus 40 Hybrid Virus

Abstract: This cell line can be a useful research tool for investigating the role of spiral ligament fibrocytes in homeostasis and inflammation of the inner ear.

Cited by 9 publications

References 43 publications

Functional interaction between mesenchymal stem cells and spiral ligament fibrocytes

Functional interaction between mesenchymal stem cells and spiral ligament fibrocytes

Toll-Like Receptor 2-Dependent NF-κB Activation Is Involved in Nontypeable Haemophilus influenzae -Induced Monocyte Chemotactic Protein 1 Up-Regulation in the Spiral Ligament Fibrocytes of the Inner Ear

Interleukin-10 Attenuates Hypochlorous Acid-Mediated Cytotoxicity to HEI-OC1 Cochlear Cells

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