Three-dimensional cultured ampullae from rats as a screening tool for vestibulotoxicity: Proof of concept using styrene

Tallandier, V.; Merlen, Lise; Chalansonnet, Monique; Boucard, S.; Thomas, Aurélie; Venet, Thomas; Pouyatos, Benoı̂t

doi:10.1016/j.tox.2023.153600

Cited by 1 publication

(2 citation statements)

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“…In the present free-floating conditions, cysts formed from the three types of vestibular epithelia, cristas, utricles and saccules. This constitutes an improvement with respect to the more frequent use of utricles alone, although at least one other example of crista culture has been published ( Tallandier et al, 2023 ). Notably, the cysts reproduced the in vivo morphology (ampullar versus macular shape) indicating that this morphology is autonomously determined by the developing vestibule and not shaped by surrounding tissues.…”

Section: Discussionmentioning

confidence: 99%

“…One interesting alternative is that of 3D cultures, in which the non-sensory epithelium surrounding the sensory patch grows until it closes a cavity that mimics the endolymphatic compartment existing in vivo . The formation of these 3D explants, named cysts, has been reported to depend on the presence of extracellular matrices, either matrigel ( Gaboyard et al, 2005 ; Brugeaud et al, 2007 ; Bartolami et al, 2011 ; Tallandier et al, 2021 ; Tallandier et al, 2023 ) or collagen-based gels ( Gnedeva et al, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

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Maturation of type I and type II rat vestibular hair cells in vivo and in vitro

Borrajo,

Sedano,

Palou

et al. 2024

Front. Cell Dev. Biol.

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Vestibular sensory epithelia contain type I and type II sensory hair cells (HCI and HCII). Recent studies have revealed molecular markers for the identification of these cells, but the precise composition of each vestibular epithelium (saccule, utricle, lateral crista, anterior crista, posterior crista) and their postnatal maturation have not been described in detail. Moreover, in vitro methods to study this maturation are not well developed. We obtained total HCI and HCII counts in adult rats and studied the maturation of the epithelia from birth (P0) to postnatal day 28 (P28). Adult vestibular epithelia hair cells were found to comprise ∼65% HCI expressing osteopontin and PMCA2, ∼30% HCII expressing calretinin, and ∼4% HCII expressing SOX2 but neither osteopontin nor calretinin. At birth, immature HCs express both osteopontin and calretinin. P28 epithelia showed an almost adult-like composition but still contained 1.3% of immature HCs. In addition, we obtained free-floating 3D cultures of the epithelia at P1, which formed a fluid-filled cyst, and studied their survival and maturation in vitro up to day 28 (28 DIV). These cultures showed good HC resiliency and maturation. Using an enriched medium for the initial 4 days, a HCI/calretinin+-HCII ratio close to the in vivo ratio was obtained. These cultures are suitable to study HC maturation and mature HCs in pharmacological, toxicological and molecular research.

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Section: Discussionmentioning

confidence: 99%