Induction of myelinating oligodendrocytes in human cortical spheroids

Abstract

Cerebral organoids provide an accessible system for investigations of cellular composition, interactions, and organization but have lacked oligodendrocytes, the myelinating glia of the central nervous system. Here we reproducibly generated oligodendrocytes and myelin in 'oligocortical spheroids' derived from human pluripotent stem cells. Molecular features consistent with those of maturing oligodendrocytes and early myelin appeared by week 20 in culture, with further maturation and myelin compaction evident by week 30. Promyelinating drugs enhanced the rate and extent of oligodendrocyte generation and myelination, and spheroids generated from human subjects with a genetic myelin disorder recapitulated human disease phenotypes. Oligocortical spheroids provide a versatile platform for studies of myelination of the developing central nervous system and offer new opportunities for disease modeling and therapeutic development.

Stem cell derived oligodendrocytes to study myelin diseases

“…Organoids, which are 3D multicellular aggregates derived from stem cells that differentiate and self‐organize are a further step into the direction of the development of a human brain model in a dish. Recently, the first oligodendrocytes enriched brain organoids have been described (Madhavan et al, ; Marton et al, ). After 20–30 weeks in culture the transcription factor MYRF, PLP1, or MBP positive oligodendrocytes have been detected, additionally wrapping of oligodendroglial processes around axons has been observed (Madhavan et al, ).…”

“…Recently, the first oligodendrocytes enriched brain organoids have been described (Madhavan et al, ; Marton et al, ). After 20–30 weeks in culture the transcription factor MYRF, PLP1, or MBP positive oligodendrocytes have been detected, additionally wrapping of oligodendroglial processes around axons has been observed (Madhavan et al, ). Using iPSC‐derived oligodendrocytes from patients with different PMD mutations, oligodendroglial phenotypes identified in 2D cultures could be recapitulated also in the 3D system demonstrating that oligocortical organoids may be suitable for in vitro modeling of myelin diseases, such as leukodystrophies (Elitt, Barbar, & Tesar, ).…”

“…Using iPSC‐derived oligodendrocytes from patients with different PMD mutations, oligodendroglial phenotypes identified in 2D cultures could be recapitulated also in the 3D system demonstrating that oligocortical organoids may be suitable for in vitro modeling of myelin diseases, such as leukodystrophies (Elitt, Barbar, & Tesar, ). Addition of oligodendroglial differentiation promoting compounds, such as T3, clemastine or ketoconazole increased the numbers of MYRF positive oligodendrocytes demonstrating that organoids can be utilized for the identification of potential remyelination promoting compounds (Madhavan et al, ). However, the long culture periods and the frequently described variability between individual organoids are substantial obstacles for the use of organoids for high throughput screens.…”

Xeno‐free culture for generation of forebrain oligodendrocyte precursor cells from human pluripotent stem cells

“…NSC induction with Laminin 511-E8 yielded a higher number of cells within 15 days of culture, hence increasing the efficiency of large-scale production ( Figure 1a). Several methods have been reported to generate OPCs without retinoic acid in order to avoid the brain stem or spinal fate (Madhavan et al, 2018;Major et al, 2017;Piao et al, 2015;Stacpoole et al, 2013;Sundberg, Skottman, Suuronen, & Narkilahti, 2010). Derivation of the forebrain fate from hPSCs is associated with a low WNT signal (Imaizumi et al, 2015).…”

“…Human pluripotent stem cells (hPSCs) can serve as an unlimited and scalable source of OPCs for cell therapy. Moreover, the discovery of induced pluripotent stem cells (iPSCs) has enabled a new era of patient‐specific disease modeling and drug screening (Douvaras et al, ; Goldman & Kuypers, ; Madhavan et al, ; Windrem et al, ). Over decades, numerous advances have improved the generation of OPCs from hPSCs (Douvaras, ; Hu, Du, Li, Ayala, & Zhang, ; Rodrigues et al, ; Stacpoole et al, ; Wang et al, ) and a related clinical trial for spinal cord injury (SCI) has been initiated (Lebkowski, ; Priest, Manley, Denham, Wirth, & Lebkowski, ).…”