Culture of organoids with vestibular cell-derived factors promotes differentiation of embryonic stem cells into inner ear vestibular hair cells

Daisuke Osaki; Yukiteru Ouji; Masaharu Sakagami; Tomotaka Kitamura; Masayasu Misu; Tadashi Kitahara; Masahide Yoshikawa

doi:10.1016/j.jbiosc.2022.11.005

Culture of organoids with vestibular cell-derived factors promotes differentiation of embryonic stem cells into inner ear vestibular hair cells

J Biosci Bioeng. 2023 Feb;135(2):143-150. doi: 10.1016/j.jbiosc.2022.11.005. Epub 2022 Dec 8.

Authors

Daisuke Osaki¹, Yukiteru Ouji², Masaharu Sakagami³, Tomotaka Kitamura⁴, Masayasu Misu⁵, Tadashi Kitahara⁶, Masahide Yoshikawa⁷

Affiliations

¹ Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan; Department of Otolaryngology - Head and Neck Surgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: dosaki@naramed-u.ac.jp.
² Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: oujix@naramed-u.ac.jp.
³ Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan; Department of Otolaryngology - Head and Neck Surgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: m.sakagami@naramed-u.ac.jp.
⁴ Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: tomotaka.k@naramed-u.ac.jp.
⁵ Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: masayasu-m@naramed-u.ac.jp.
⁶ Department of Otolaryngology - Head and Neck Surgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: tkitahara@naramed-u.ac.jp.
⁷ Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Electronic address: myoshika@naramed-u.ac.jp.

PMID: 36503871
DOI: 10.1016/j.jbiosc.2022.11.005

Abstract

Vestibular hair cells (V-HCs) residing in the inner ear have important roles related to balance. Although differentiation of pluripotent stem cells into HCs has been shown, an effective method has yet to be established. We previously reported that use of vestibular cell-derived conditioned medium (V-CM) was helpful to induce embryonic stem (ES) cells to differentiate into V-HC-like cells in two-dimensional (2D) cultures of ES-derived embryoid bodies (EBs). In the present report, V-CM was used with three-dimensional (3D) cultures of EBs, which resulted in augmented expression of V-HC-related markers (Math1, Myosin6, Brn3c, Dnah5), but not of the cochlear HC-related marker Lmod3. Gene expression analyses of both 2D and 3D EBs cultured for two weeks revealed a greater level of augmented induction of HC-related markers in the 3D-cultured EBs. These results indicate that a 3D culture in combination with use of V-CM is an effective method for producing V-HCs.

Keywords: Conditioned medium; Differentiation; Embryonic stem cells; Hair cells; Organoid; Vestibular.

MeSH terms

Cell Differentiation / genetics
Cells, Cultured
Embryonic Stem Cells
Hair Cells, Auditory, Inner / metabolism
Hair Cells, Vestibular*
Organoids