PMID- 26003451 OWN - NLM STAT- MEDLINE DCOM- 20151016 LR - 20150620 IS - 1872-7972 (Electronic) IS - 0304-3940 (Linking) VI - 599 DP - 2015 Jul 10 TI - Limited hair cell induction from human induced pluripotent stem cells using a simple stepwise method. PG - 49-54 LID - S0304-3940(15)00389-4 [pii] LID - 10.1016/j.neulet.2015.05.032 [doi] AB - Disease-specific induced pluripotent stem cells (iPS) cells are expected to contribute to exploring useful tools for studying the pathophysiology of inner ear diseases and to drug discovery for treating inner ear diseases. For this purpose, stable induction methods for the differentiation of human iPS cells into inner ear hair cells are required. In the present study, we examined the efficacy of a simple induction method for inducing the differentiation of human iPS cells into hair cells. The induction of inner ear hair cell-like cells was performed using a stepwise method mimicking inner ear development. Human iPS cells were sequentially transformed into the preplacodal ectoderm, otic placode, and hair cell-like cells. As a first step, preplacodal ectoderm induction, human iPS cells were seeded on a Matrigel-coated plate and cultured in a serum free N2/B27 medium for 8 days according to a previous study that demonstrated spontaneous differentiation of human ES cells into the preplacodal ectoderm. As the second step, the cells after preplacodal ectoderm induction were treated with basic fibroblast growth factor (bFGF) for induction of differentiation into otic-placode-like cells for 15 days. As the final step, cultured cells were incubated in a serum free medium containing Matrigel for 48 days. After preplacodal ectoderm induction, over 90% of cultured cells expressed the genes that express in preplacodal ectoderm. By culture with bFGF, otic placode marker-positive cells were obtained, although their number was limited. Further 48-day culture in serum free media resulted in the induction of hair cell-like cells, which expressed a hair cell marker and had stereocilia bundle-like constructions on their apical surface. Our results indicate that hair cell-like cells are induced from human iPS cells using a simple stepwise method with only bFGF, without the use of xenogeneic cells. CI - Copyright (c) 2015 Elsevier Ireland Ltd. All rights reserved. FAU - Ohnishi, Hiroe AU - Ohnishi H AD - Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. FAU - Skerleva, Desislava AU - Skerleva D AD - Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. FAU - Kitajiri, Shin-ichiro AU - Kitajiri S AD - Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. FAU - Sakamoto, Tatsunori AU - Sakamoto T AD - Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. FAU - Yamamoto, Norio AU - Yamamoto N AD - Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. FAU - Ito, Juichi AU - Ito J AD - Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. FAU - Nakagawa, Takayuki AU - Nakagawa T AD - Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. Electronic address: tnakagawa@ent.kuhp.kyoto-u.ac.jp. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150521 PL - Ireland TA - Neurosci Lett JT - Neuroscience letters JID - 7600130 RN - 0 (Culture Media, Serum-Free) RN - 103107-01-3 (Fibroblast Growth Factor 2) SB - IM MH - Cell Differentiation MH - Cells, Cultured MH - Culture Media, Serum-Free MH - Ectoderm/cytology MH - Fibroblast Growth Factor 2/pharmacology MH - Hair Cells, Auditory, Inner/*cytology MH - Humans MH - Induced Pluripotent Stem Cells/*cytology/drug effects OTO - NOTNLM OT - Disease-specific iPS cells OT - Inner ear hair cells OT - Otic placode OT - Pluripotent stem cell OT - Preplacodal ectoderm OT - bFGF EDAT- 2015/05/25 06:00 MHDA- 2015/10/17 06:00 CRDT- 2015/05/25 06:00 PHST- 2015/02/19 00:00 [received] PHST- 2015/05/16 00:00 [revised] PHST- 2015/05/18 00:00 [accepted] PHST- 2015/05/25 06:00 [entrez] PHST- 2015/05/25 06:00 [pubmed] PHST- 2015/10/17 06:00 [medline] AID - S0304-3940(15)00389-4 [pii] AID - 10.1016/j.neulet.2015.05.032 [doi] PST - ppublish SO - Neurosci Lett. 2015 Jul 10;599:49-54. doi: 10.1016/j.neulet.2015.05.032. Epub 2015 May 21.