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as a potential tool to treat sensorineural hearing loss by aiding the regeneration of spiral ganglion neurons (SGNs) in the inner ear. However, transplantation procedures have yet to be explored thoroughly to ensure proper cell differentiation and optimal transplant procedures. We hypothesized that the aggregation of human embryonic stem cell (hESC)-derived otic neuronal progenitor (ONP) cells into a multicellular form would improve their function and their survivalin vivopost-transplantation. We generated hESC-derived ONP spheroids-an aggregate form conducive to differentiation, transplantation, and prolonged cell survival-to optimize conditions for their transplantation. Our findings indicate that these cell spheroids maintain the molecular and functional characteristics similar to those of ONP cells, which are upstream in the SGN lineage. Moreover, our phenotypical, electrophysiological, and mechanical data suggest an optimal spheroid transplantation point after 7 days ofin vitrothree-dimensional (3D) culture. We have also developed a feasible transplantation protocol for these spheroids using a micropipette aided by a digital microinjection system. In summary, the present work demonstrates that the transplantation of ONP cells in spheroid form into the inner ear through micropipette 7 days after seeding for 3D spheroid culture is an expedient and viable method for stem cell replacement therapies in the inner ear.Impact statement Sensorineural hearing loss affects millions of people worldwide. Although inner ear stem cell replacement therapies offer a promising method to mitigate this hearing loss by aiding in the regeneration of spiral ganglion neurons, fewin vivostudies have been performed in this area. By providing detailed cell spheroid characterization, determining an optimal development stage for transplantation, and establishing a reliable and reproducible transplantation protocol, we aimed to provide the necessary details to bridge the gap betweenin vitroexperiments andin vivostudies of stem cell replacement therapies in the inner ear.", "subjects" : [ { "otype" : "Classification", "mtid" : 10545, "link" : "/api/classification/10545", "label" : "Biológiai tudományok", "published" : true, "snippet" : true }, { "otype" : "Classification", "mtid" : 11065, "link" : "/api/classification/11065", "label" : "Ipari biotechnológia", "published" : true, "snippet" : true }, { "otype" : "Classification", "mtid" : 11047, "link" : "/api/classification/11047", "label" : "Orvostechnikai műszaki tudományok", "published" : true, "snippet" : true } ], "keywords" : [ { "otype" : "Keyword", "mtid" : 1042712, "link" : "/api/keyword/1042712", "label" : "Stem Cell Niche", "published" : true, "oldId" : 1042712, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1093637, "link" : "/api/keyword/1093637", "label" : "inner ear", "published" : true, "oldId" : 1093637, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1121680, "link" : "/api/keyword/1121680", "label" : "Human embryonic stem cells", "published" : true, "oldId" : 1121680, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1586799, "link" : "/api/keyword/1586799", "label" : "Spheroid", "published" : true, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1622406, "link" : "/api/keyword/1622406", "label" : "three-dimensional culture", "published" : true, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1742162, "link" : "/api/keyword/1742162", "label" : "Spiral ganglion neurons", "published" : true, "snippet" : true }, { "otype" : "Keyword", "mtid" : 2082565, "link" : "/api/keyword/2082565", "label" : "stem cell replacement therapy", "published" : true, "snippet" : true } ], "digital" : null, "printed" : null, "sourceYear" : 2020, "foreignEdition" : true, "foreignLanguage" : true, "fullPublication" : true, "conferencePublication" : false, "nationalOrigin" : null, "missingAuthor" : false, "oaType" : "NONE", "oaCheckDate" : "2022-10-11", "oaFree" : false, "citationCount" : 0, "citationCountUnpublished" : 0, "citationCountWoOther" : 0, "independentCitCountWoOther" : 0, "doiCitationCount" : 0, "wosCitationCount" : 0, "scopusCitationCount" : 0, "independentCitationCount" : 0, "unhandledCitationCount" : 0, "citingPubCount" : 0, "independentCitingPubCount" : 0, "unhandledCitingPubCount" : 0, "citedPubCount" : 1, "citedCount" : 1, "ratings" : [ { "otype" : "SjrRating", "mtid" : 11196279, "link" : "/api/sjrrating/11196279", "label" : "sjr:Q2 (2021) Scopus - Biochemistry TISSUE ENGINEERING PART A 1937-3341 1937-335X", "listPos" : 191, "rankValue" : 0.49, "type" : "journal", "ratingType" : { "otype" : "RatingType", "mtid" : 10002, "link" : "/api/ratingtype/10002", "label" : "sjr", "code" : "sjr", "published" : true, "snippet" : true }, "subject" : { "otype" : "ClassificationExternal", "mtid" : 1303, "link" : "/api/classificationexternal/1303", "label" : "Scopus - Biochemistry", "published" : true, "oldId" : 1303, "snippet" : true }, "ranking" : "Q2", "calculation" : "DIRECT", "published" : true, "snippet" : true } ], "ratingsForSort" : "Q2", "hasCitationDuplums" : false, "userChangeableUntil" : "2022-01-23T13:29:43.420+0000", "directInstitutesForSort" : "", "ownerAuthorCount" : 8, "ownerInstituteCount" : 76, "directInstituteCount" : 0, "authorCount" : 10, "contributorCount" : 0, "hasQualityFactor" : true, "link" : "/api/publication/31467828", "label" : "Heuer Rachel A. et al. 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