Verkerke Marloes, Berdenis van Berlekom Amber, Donega Vanessa, Vonk Daniëlle, Sluijs Jacqueline A, Butt Nayab F, Kistemaker Lois, de Witte Lot D, Pasterkamp R Jeroen, Middeldorp Jinte, Hol Elly M
Department of Translational Neuroscience, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Department of Psychiatry, University Medical Center Utrecht, Brain Center, Utrecht University, Utrecht, The Netherlands.
Glia. 2024 Feb;72(2):362-374. doi: 10.1002/glia.24479. Epub 2023 Oct 17.
Cerebral organoids (CerOrgs) derived from human induced pluripotent stem cells (iPSCs) are a valuable tool to study human astrocytes and their interaction with neurons and microglia. The timeline of astrocyte development and maturation in this model is currently unknown and this limits the value and applicability of the model. Therefore, we generated CerOrgs from three healthy individuals and assessed astrocyte maturation after 5, 11, 19, and 37 weeks in culture. At these four time points, the astrocyte lineage was isolated based on the expression of integrin subunit alpha 6 (ITGA6). Based on the transcriptome of the isolated ITGA6-positive cells, astrocyte development started between 5 and 11 weeks in culture and astrocyte maturation commenced after 11 weeks in culture. After 19 weeks in culture, the ITGA6-positive astrocytes had the highest expression of human mature astrocyte genes, and the predicted functional properties were related to brain homeostasis. After 37 weeks in culture, a subpopulation of ITGA6-negative astrocytes appeared, highlighting the heterogeneity within the astrocytes. The morphology shifted from an elongated progenitor-like morphology to the typical bushy astrocyte morphology. Based on the morphological properties, predicted functional properties, and the similarities with the human mature astrocyte transcriptome, we concluded that ITGA6-positive astrocytes have developed optimally in 19-week-old CerOrgs.
源自人类诱导多能干细胞（iPSC）的脑类器官（CerOrg）是研究人类星形胶质细胞及其与神经元和小胶质细胞相互作用的宝贵工具。目前尚不清楚该模型中星形胶质细胞发育和成熟的时间线，这限制了该模型的价值和适用性。因此，我们从三名健康个体中生成了脑类器官，并在培养5周、11周、19周和37周后评估了星形胶质细胞的成熟情况。在这四个时间点，基于整联蛋白亚基α6（ITGA6）的表达分离出星形胶质细胞谱系。根据分离出的ITGA6阳性细胞的转录组，星形胶质细胞发育在培养5至11周之间开始，星形胶质细胞成熟在培养11周后开始。培养19周后，ITGA6阳性星形胶质细胞具有人类成熟星形胶质细胞基因的最高表达，并且预测的功能特性与脑稳态相关。培养37周后，出现了ITGA6阴性星形胶质细胞亚群，突出了星形胶质细胞内的异质性。形态从细长的祖细胞样形态转变为典型的浓密星形胶质细胞形态。基于形态学特性、预测的功能特性以及与人类成熟星形胶质细胞转录组的相似性，我们得出结论，ITGA6阳性星形胶质细胞在19周龄的脑类器官中已最佳发育。
