Nadadhur Aishwarya G, Leferink Prisca S, Holmes Dwayne, Hinz Lisa, Cornelissen-Steijger Paulien, Gasparotto Lisa, Heine Vivi M
Department of Functional Genomics, Center for Neurogenomics and Cognitive Research, Amsterdam Neuroscience, Vrije Universiteit Amsterdam, the Netherlands.
Pediatric Neurology, Emma Children's Hospital, Amsterdam UMC, Amsterdam Neuroscience, Vrije Universiteit Amsterdam, the Netherlands.
Stem Cell Res. 2018 Oct;32:25-34. doi: 10.1016/j.scr.2018.08.017. Epub 2018 Aug 23.
The neural tube consists of neural progenitors (NPs) that acquire different characteristics during gestation due to patterning factors. However, the influence of such patterning factors on human pluripotent stem cells (hPSCs) during in vitro neural differentiation is often unclear. This study compared neural induction protocols involving in vitro patterning with single SMAD inhibition (SSI), retinoic acid (RA) administration and dual SMAD inhibition (DSI). While the derived NP cells expressed known NP markers, they differed in their NP expression profile and differentiation potential. Cortical neuronal cells generated from 1) SSI NPs exhibited less mature neuronal phenotypes, 2) RA NPs exhibited an increased GABAergic phenotype, and 3) DSI NPs exhibited greater expression of glutamatergic lineage markers. Further, although all NPs generated astrocytes, astrocytes derived from the RA-induced NPs had the highest GFAP expression. Differences between NP populations included differential expression of regional identity markers HOXB4, LBX1, OTX1 and GSX2, which persisted into mature neural cell stages. This study suggests that patterning factors regulate how potential NPs may differentiate into specific neuronal and glial cell types in vitro. This challenges the utility of generic neural induction procedures, while highlighting the importance of carefully selecting specific NP protocols.
神经管由神经祖细胞（NP）组成，这些神经祖细胞在妊娠期由于模式形成因子而获得不同的特征。然而，在体外神经分化过程中，此类模式形成因子对人类多能干细胞（hPSC）的影响往往并不明确。本研究比较了涉及体外模式形成的神经诱导方案与单一SMAD抑制（SSI）、视黄酸（RA）给药和双重SMAD抑制（DSI）。虽然所产生的NP细胞表达已知的NP标志物，但它们在NP表达谱和分化潜能方面存在差异。从1）SSI NP产生的皮质神经元细胞表现出不太成熟的神经元表型，2）RA NP表现出增加的GABA能表型，3）DSI NP表现出更高的谷氨酸能谱系标志物表达。此外，虽然所有NP都产生了星形胶质细胞，但源自RA诱导的NP的星形胶质细胞具有最高的GFAP表达。NP群体之间的差异包括区域身份标志物HOXB4、LBX1、OTX1和GSX2的差异表达，这些差异一直持续到成熟神经细胞阶段。本研究表明，模式形成因子调节潜在的NP在体外如何分化为特定的神经元和胶质细胞类型。这对通用神经诱导程序的实用性提出了挑战，同时强调了仔细选择特定NP方案的重要性。
