02127nas a2200361   4500000000100000008004100001260001500042653002000057653001500077653002300092653001400115653002400129653002000153100001700173700002000190700002600210700001900236700001200255700001500267700002100282700001400303700001900317700001300336700002300349700001800372700002100390245008700411856007200498300001700570490000700587520115700594022001401751       2024                            d  c2024-12-0510aPurkinje neuron10amorphogens10amultiplexed screen10aorganoids10asingle-cell RNA-seq10atransplantation1 aNeal D. Amin1 aKevin W. Kelley1 aKonstantin Kaganovsky1 aMassimo Onesto1 aJin Hao1 aYuki Miura1 aJames P. McQueen1 aNoah Reis1 aGenta Narazaki1 aTommy Li1 aShravanti Kulkarni1 aSergey Pavlov1 aSergiu P. Pașca00aGenerating human neural diversity with a multiplexed morphogen screen in organoids  uhttps://www.sciencedirect.com/science/article/pii/S1934590924003783  a1831-1846.e90 v313 aMorphogens choreograph the generation of remarkable cellular diversity in the developing nervous system. Differentiation of stem cells in vitro often relies upon the combinatorial modulation of these signaling pathways. However, the lack of a systematic approach to understand morphogen-directed differentiation has precluded the generation of many neural cell populations, and the general principles of regional specification and maturation remain incomplete. Here, we developed an arrayed screen of 14 morphogen modulators in human neural organoids cultured for over 70 days. Deconvolution of single-cell-multiplexed RNA sequencing data revealed design principles of brain region specification. We tuned neural subtype diversity to generate a tachykinin 3 (TAC3)-expressing striatal interneuron type within assembloids. To circumvent limitations of in vitro neuronal maturation, we used a neonatal rat transplantation strategy that enabled human Purkinje neurons to develop their hallmark complex dendritic branching. This comprehensive platform yields insights into the factors influencing stem cell-derived neural diversification and maturation.  a1934-5909