The effects of substrate composition and topography on the characteristics and growth of cell cultures of cochlear fibrocytes

Mahendrasingam, Shanthini; Osborn, Anya Helen; Israr, Muhammad Zubair; Riasat, Kiran; Gater, Rachel; Spanò, Andrea; Gogo-Fyneface, Desire; Worrall, Amy J; Furness, David N.

doi:10.1016/j.heares.2021.108427

Cited by 1 publication

(3 citation statements)

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“…After initial seeding, the fibrocytes appeared rounded with no processes or connections. As time progressed, the cells became more interconnected and less flat than those cultured on 2-D collagen-coated surfaces (see Mahendrasingam et al 2022 ). Cells were examined for morphological changes over time ( n = 5 runs), Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The fact that cultured fibrocytes labelled for both markers has prevented us from a definitive identification of fibrocyte type (Mahendrasingam et al 2011a ). This might be related to the culture conditions or to the immature state of the fibrocytes (Mahendrasingam et al 2022 ). One possibility is that they most closely resemble type III fibrocytes (since they label for aquaporin), with some upregulation of S-100 presumably as an adaptation to the absence of other fibrocyte types (Wu and Marcus 2003 ; Mahendrasingam et al 2011a ).…”

Section: Discussionmentioning

confidence: 99%

“…We have grown cochlear fibrocytes on collagen I gels in order to assess their electrical properties and their labelling for key fibrocyte marker proteins. We chose to culture the cells on collagen hydrogels, rather than on flat surfaces, since this approach produces fibrocytes exhibiting a more natural morphology (Mahendrasingam et al 2022 ). We also compared their electrical properties with those of fibrocytes within the lateral wall, either in dissected portions or in cochlear slices.…”

Section: Introductionmentioning

confidence: 99%

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Electrical and Immunohistochemical Properties of Cochlear Fibrocytes in 3D Cell Culture and in the Excised Spiral Ligament of Mice

Osborn

Caruana

Furness

et al. 2022

JARO

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Fibrocyte degeneration in the cochlear lateral wall is one possible pathology of age-related metabolic hearing loss (presbycusis). Within the lateral wall fibrocytes play a role in potassium recycling and maintenance of the endocochlear potential. It has been proposed that cell replacement therapy could prevent fibrocyte degeneration in the CD/1 mouse model of hearing loss. For this to work, the replacement fibrocytes would need to take over the structural and physiological role of those lost. We have grown lateral wall fibrocytes from neonatal CD/1 mice in a 3D-collagen gel culture with the aim of assessing their functional similarity to native lateral wall fibrocytes, the latter in a slice preparation and in excised spiral ligament pieces. We have compared cultured and native fibrocytes using both immuno-labelling of characteristic proteins and single cell electrophysiology. Cultured fibrocytes exhibited rounded cell bodies with extending processes. They labelled with marker antibodies targeting aquaporin 1 and calcium-binding protein S-100, precluding an unambiguous identification of fibrocyte type. In whole-cell voltage clamp, both native and cultured fibrocytes exhibited non-specific currents and voltage-dependent K+ currents. The non-specific currents from gel-cultured and excised spiral ligament fibrocytes were partially and reversibly blocked by external TEA (10 mM). The TEA-sensitive current had a mean reversal potential of + 26 mV, suggesting a permeability sequence of Na+ > K+. These findings indicate that 3D-cultured fibrocytes share a number of characteristics with native spiral ligament fibrocytes and thus might represent a suitable population for transplantation therapy aimed at treating age-related hearing loss.

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Section: Resultsmentioning

confidence: 99%