Six-month cultured cerebral organoids from human ES cells contain matured neural cells

Abstract

Recently, researchers have developed protocols for human cerebral organoids using human pluripotent stem cells, which mimic the structure of the developing human brain. Existing research demonstrated that human cerebral organoids which undergo short cultivation periods, contain astrocytes, neurons, and neural stem cells, but lacked mature oligodendrocytes, and mature, fully functional neurons. In this study, we analyzed organoids induced from H9 human embryonic stem (ES) cells that were cultivated for as long as six months. We observed mature oligodendrocytes, positive for MBP (myelin-basic protein), and mature GAD67 (glutamate decarboxylase 67 kDa isoform)-positive inhibitory neurons and VGLUT1 (vesicular glutamate transporter 1)-positive excitatory neurons via immunohistochemical analysis. These observations suggest that long-term cultivation of cerebral organoids can lead to the maturation of human cerebral organoids, which can be used as a tool to study the development of human brains.

Vascularization and Engraftment of Transplanted Human Cerebral Organoids in Mouse Cortex

“…This may reflect stronger glial differentiation signals at the transplant site in vivo than in cultured organoids. Concordantly, as compared to in vitro cerebral organoid cultures where oligodendrocytes were detected only at later stages ( Renner et al, 2017 ; Matsui et al, 2018 ), we detected ∼10% donor cells expressing Olig2 at both two and four weeks after grafting. Of note, during human neurodevelopment, Olig2 is expressed in OPC but also in a subset of neuroprogenitors ( Jakovcevski and Zecevic, 2005 ), thus the Olig2+ cells in the organoid grafts may represent both populations.…”

Dynamic Characterization of Structural, Molecular, and Electrophysiological Phenotypes of Human-Induced Pluripotent Stem Cell-Derived Cerebral Organoids, and Comparison with Fetal and Adult Gene Profiles

“…Moving forward from the iPSC stage, we noted a downregulation in the OCT4 pluripotency marker and Pax6 neural stem cell marker, and an upregulation in the markers of neural cell types (neurons and astrocytes; Figure 2). In a similar manner to the current study, others recently reported a similar upregulation in neurons and glia as cerebral organoid differentiation from stem cells progressed [22,23]. Interestingly, we also observed what appeared to be an attempt by the cerebral organoids to produce blood vessels (CD31 and SMA), (Figure 2), suggesting the presence of cell types not originating from the ectoderm [24].…”

Transplantation of Human Brain Organoids: Revisiting the Science and Ethics of Brain Chimeras

“…Subsequent studies refined these aspects of normal neurodevelopment in region-specific organoids, resulting in rudimentary segregation of superficial and deep cortical layers (Paşca et al, 2015;Qian et al, 2016), generation of a distinct layer of outer radial glial cells (Qian et al, 2016), and expansion of cortical folds (Li et al, 2017). Glial populations were observed at later time points, including astrocytes (Dezonne et al, 2017;Paşca et al, 2015;Qian et al, 2016;Sloan et al, 2017) and oligodendrocytes (Matsui et al, 2018). Integration of interneurons into these cortical organoids has been studied through fusion of dorsalized glutamatergic organoids with ventralized organoids containing GABAergic neurons (Bagley et al, 2017;Birey et al, 2017;Xiang et al, 2017).…”