Recapitulating cortical development with organoid culture in vitro and modeling abnormal spindle-like (ASPM related primary) microcephaly disease

Abstract

The development of a cerebral organoid culture in vitro offers an opportunity to generate human brain-like organs to investigate mechanisms of human disease that are specific to the neurogenesis of radial glial (RG) and outer radial glial (oRG) cells in the ventricular zone (VZ) and subventricular zone (SVZ) of the developing neocortex. Modeling neuronal progenitors and the organization that produces mature subcortical neuron subtypes during early stages of development is essential for studying human brain developmental diseases. Several previous efforts have shown to grow neural organoid in culture dishes successfully, however we demonstrate a new paradigm that recapitulates neocortical development process with VZ, OSVZ formation and the lamination organization of cortical layer structure. In addition, using patient-specific induced pluripotent stem cells (iPSCs) with dysfunction of the Aspm gene from a primary microcephaly patient, we demonstrate neurogenesis defects result in defective neuronal activity in patient organoids, suggesting a new strategy to study human developmental diseases in central nerve system.Electronic supplementary materialThe online version of this article (doi:10.1007/s13238-017-0479-2) contains supplementary material, which is available to authorized users.

Vascularized human cortical organoids model cortical development in vivo

“…Several previous studies have illustrated that neurons in cortical organoids without vascular structures are ultimately able to reach mature states as their morphology and functionality progressively mature (Lancaster et al, 2013;Li et al, 2017;Qian et al, 2016). Since vOrganoids partially resemble human cortical development in their molecular and cell subtype aspects, we next investigated the functional characteristics of the neurons with electrophysiological recording.…”

“…In our previous studies, we successfully established an appropriate approach to generate cerebral organoids from hESCs (human embryonic stem cells) or hiPSCs that can recapitulate in vivo human cortical development with the formation of a well-polarized ventricle neuroepithelial structure that consists of vRG, oRG and IPC cells and the production of mature neurons within layers (Li et al, 2017). However, a major limitation in the current culture scheme that is preventing a truly in-vivo-like functionality has been the lack of a microenvironment, such as the vascular circulation.…”

Using brain organoids to study human neurodevelopment, evolution and disease

“…Abbreviations: IP, intermediate progenitor; a/bRG, apical/basal radial glial cell; sc-RNA-seq, single cell RNA sequencing process, the functional aspects and physiological properties of the human neurons when found in a 3D structure. Toward this end, electrophysiological recording and calcium imaging methodologies have been developed to measure the neuron's capability of firing action potentials (Bershteyn et al, 2017;Li, Sun, et al, 2017;Qian et al, 2016;Quadrato et al, 2017;Watanabe et al, 2017), or to measure the response of the neuronal networks to physiological sensory stimuli by applying optogenetics (Quadrato et al, 2017) (Figure 3d). Nonetheless, the lack of organization of multiple brain regions within the cerebral organoids limits the study of neuronal connectivity -a fundamental feature of the human brain.…”

Stem Cells, Genome Editing, and the Path to Translational Medicine

“…Organoid models are now a common tool to study neurodevelopmental disorders, including microcephaly (Lancaster et al, 2013; Li et al, 2017a), autism spectrum disease associated with macrocephaly (Mariani et al, 2017; Wang et al, 2017), lissencephally (Bershteyn et al, 2017; Karzbrun et al, 2018), Rett Syndrom (Mellios et al, 2017), Thymothy syndrome (Birey et al, 2017), lysosomal storage diseases such as Sandhoff disease (Allende et al, 2018), schizophrenia (Srikanth et al, 2018; Stachowiak et al, 2017) and Zika infection on neonatal brain development (reviewed in (Qian et al, 2017). Despite the early success with this approach for disease-modelling, challenges remain.…”