1. Lgr5+ endogenous progenitor cells in the adult (deafened) cochlea.
- Author
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Smith-Cortinez, N., Holtappels, F., Fenton, G., Versnel, H., Thomeer, H. G. X. M., and Straatman, L. V.
- Subjects
CONFERENCES & conventions ,INNER ear ,DEAFNESS ,STEM cells ,COCHLEA ,HAIR cells - Abstract
Introduction: LGR5 positive supporting cells (SCs) in the cochlea give rise to hair cells (HCs) during embryonic development. Neonatal SCs have increased progenitor potential compared to adult and only a few studies (including ours) have shown survival of SCs with progenitor cell markers after severe HC loss in adult mice. In mammals, there is no evidence for spontaneous HC regeneration in adulthood. However, three-dimensional cultures have allowed the expansion and experimentation of human (and mouse) inner ear organoids. Here, we evaluated HC differentiation from human cochlear organoids and from adult normal-hearing and deafened mice. Material and methods: Adult patients undergoing surgery for skull base tumors were included. Sensory epithelium of the cochlea and vestibular organ was collected in medium and tissue was digested to single cell suspension. Adult Lgr5-eGFP-IRES-creERT2 heterozygous mice were used. Mice were deafened with a single dose of furosemide in combination with kanamycin and deafening was confirmed by auditory brainstem responses (ABRs). Cochleas were harvested and digested to single cell suspension and after filtering, 3D drops were made with Matrigel. Cells were grown on expansion medium (EM) for 10 days and differentiation medium (DM) for 3-10 days after. Organoids were fixed, permea-bilized and processed for immunofluorescence and whole-mounted for imaging in a confocal microscope. Results: Vestibular-organ-derived organoids were generated in EM from all seven patients so far included. Cochlea-derived organoids were generated in five out of seven patients. After exposure to DM, vestibular organ-derived and cochlea-derived organoids produced MYO7A+ HC-like cells. Cochlear organoids from normal-hearing mice expressed LGR5 and Ki67 in EM and MYO7A after differentiation. Significantly less cochlear organoids were produced from deafened mice; however the organoids reached similar size as NH-cochlear organoids, expressed LGR5 and Ki67 in EM and MYO7A after differentiation. Conclusions: Cochlear and vestibular tissue from adult patients (and adult normal-hearing and deafened mice) possess progenitor potential and the capacity to generate inner ear organoids in vitro. After differentiation, HCs were visible in tissue derived from human cochlea, human vestibular organ, and adult mouse cochlea. The adult inner ear has (limited) regenerative capacity and can produce new MYO7A+ HCs. [ABSTRACT FROM AUTHOR]
- Published
- 2024