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Dysregulation of their development underlies movement disorders and medulloblastomas. It is suspected that these disorders arise in progenitor states of the CGN lineage, for which human models are lacking. Here, we have differentiated human hindbrain neuroepithelial stem (hbNES) cells to CGNs in vitro using soluble growth factors, recapitulating key progenitor states in the lineage. We show that hbNES cells are not lineage committed and retain rhombomere 1 regional identity. Upon differentiation, hbNES cells transit through a rhombic lip (RL) progenitor state at day 7, demonstrating human specific sub-ventricular cell identities. This RL state is followed by an ATOH1+ CGN progenitor state at day 14. By the end of a 56-day differentiation procedure, we obtain functional neurons expressing CGN markers GABAAR\u03b16 and vGLUT2. We show that sonic hedgehog promotes GABAergic lineage specification and CGN progenitor proliferation. Our work presents a new model with which to study development and diseases of the CGN lineage in a human context.<\/jats:p>","DOI":"10.1242\/dev.201534","type":"journal-article","created":{"date-parts":[[2023,6,29]],"date-time":"2023-06-29T04:04:34Z","timestamp":1688011474000},"source":"Crossref","is-referenced-by-count":0,"title":["Directed differentiation of human hindbrain neuroepithelial stem cells recapitulates cerebellar granule neurogenesis"],"prefix":"10.1242","volume":"150","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1072-1886","authenticated-orcid":false,"given":"Biren M.","family":"Dave","sequence":"first","affiliation":[{"name":"Temerty Faculty of Medicine, University of Toronto 1 Department of Molecular Genetics , , Toronto, ON M5S1A8 , Canada"},{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xin","family":"Chen","sequence":"additional","affiliation":[{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"},{"name":"Songjiang Research Institute, Songjiang Hospital, Shanghai Jiao Tong University School of Medicine 3 , Shanghai 201600 , China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fraser","family":"McCready","sequence":"additional","affiliation":[{"name":"Temerty Faculty of Medicine, University of Toronto 1 Department of Molecular Genetics , , Toronto, ON M5S1A8 , Canada"},{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Connor S.","family":"Charton","sequence":"additional","affiliation":[{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"},{"name":"University of Toronto 4 Department of Laboratory Medicine and Pathobiology, Temerty Faculty of Medicine , , Toronto, ON M5S1A8 , Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rachel M.","family":"Morley","sequence":"additional","affiliation":[{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"},{"name":"Edinburgh Medical School, The University of Edinburgh 5 , Edinburgh EH16 4SB , UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jignesh K.","family":"Tailor","sequence":"additional","affiliation":[{"name":"Riley Hospital for Children 6 Department of Neurosurgery , , Indianapolis, IN 46202 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4400-0091","authenticated-orcid":false,"given":"James","family":"Ellis","sequence":"additional","affiliation":[{"name":"Temerty Faculty of Medicine, University of Toronto 1 Department of Molecular Genetics , , Toronto, ON M5S1A8 , Canada"},{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xi","family":"Huang","sequence":"additional","affiliation":[{"name":"Temerty Faculty of Medicine, University of Toronto 1 Department of Molecular Genetics , , Toronto, ON M5S1A8 , Canada"},{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5718-6465","authenticated-orcid":false,"given":"Peter B.","family":"Dirks","sequence":"additional","affiliation":[{"name":"Temerty Faculty of Medicine, University of Toronto 1 Department of Molecular Genetics , , Toronto, ON M5S1A8 , Canada"},{"name":"The Hospital for Sick Children 2 Developmental and Stem Cell Biology Program , , Toronto, ON M5G0A4 , Canada"},{"name":"University of Toronto 4 Department of Laboratory Medicine and Pathobiology, Temerty Faculty of Medicine , , Toronto, ON M5S1A8 , Canada"},{"name":"The Hospital for Sick Children 7 Division of Neurosurgery , , Toronto, ON M5G1X8 , Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"237","published-online":{"date-parts":[[2023,7,7]]},"reference":[{"key":"2023071807252855800_DEV201534C1","doi-asserted-by":"publisher","first-page":"8730","DOI":"10.1074\/jbc.270.15.8730","article-title":"A mammalian helix-loop-helix factor structurally related to the product of Drosophila proneural gene atonal is a positive transcriptional regulator expressed in the developing nervous system","volume":"270","author":"Akazawa","year":"1995","journal-title":"J. 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