Chang So-Young, Carpena Nathaniel T, Mun Seyoung, Jung Jae Yun, Chung Phil-Sang, Shim Hosup, Han Kyudong, Ahn Jin-Chul, Lee Min Young
Beckman Laser Institute Korea, Dankook University, 119 Dandae-ro, Cheonan 31116, Republic of Korea.
Department of Otolaryngology-Head & Neck Surgery, College of Medicine, Dankook University, 119 Dandae-ro, Cheonan 31116, Republic of Korea.
Mol Ther Methods Clin Dev. 2020 Mar 13;17:556-567. doi: 10.1016/j.omtm.2020.03.010. eCollection 2020 Jun 12.
Photobiomodulation (PBM) stimulates different types of stem cells to migrate, proliferate, and differentiate and . However, little is known about the effects of PBM on the differentiation of embryonic stem cells (ESCs) toward the otic lineage. Only a few reports have documented the differentiation of ESCs into inner-ear hair cells (HCs) due to the complexity of HCs compared with other target cell types. In this study, we determined the optimal condition to differentiate the ESCs into the otic organoid using different culture techniques and PBM parameters. The efficiency of organoid formation within the embryoid body (EB) was dependent on the cell density of the hanging drop. PBM, using 630 nm wavelength light-emitting diodes (LEDs), further improved the differentiation of inner-ear hair cell-like cells coupled with reactive oxygen species (ROS) overexpression. Transcriptome analysis showed the factors that are responsible for the effect of PBM in the formation of otic organoids, notably, the downregulation of neural development-associated genes and the hairy and enhancer of split 5 () gene, which inhibits the differentiation of prosensory cells to hair cells. These data enrich the current differentiation protocols for generating inner-ear hair cells.
光生物调节（PBM）可刺激不同类型的干细胞迁移、增殖和分化。然而，关于PBM对胚胎干细胞（ESC）向耳系分化的影响知之甚少。由于与其他靶细胞类型相比，内耳毛细胞（HC）的复杂性，仅有少数报告记录了ESC向内耳毛细胞的分化。在本研究中，我们使用不同的培养技术和PBM参数确定了将ESC分化为耳类器官的最佳条件。胚状体（EB）内类器官形成的效率取决于悬滴的细胞密度。使用630nm波长发光二极管（LED）的PBM进一步改善了内耳毛细胞样细胞的分化，并伴有活性氧（ROS）的过表达。转录组分析显示了PBM在耳类器官形成中起作用的相关因子，特别是与神经发育相关基因和抑制前感觉细胞向毛细胞分化的分裂增强子5（hairy and enhancer of split 5，HES5）基因的下调。这些数据丰富了目前生成内耳毛细胞的分化方案。
