Cerebral organoids recapitulate human brain development at a considerable level of detail, even in the absence of externally added signaling factors. The patterning events driving this self-organization are currently unknown. Here, we examine the developmental and differentiative capacity of cerebral organoids. Focusing on forebrain regions, we demonstrate the presence of a variety of discrete ventral and dorsal regions. Clearing and subsequent 3D reconstruction of entire organoids reveal that many of these regions are interconnected, suggesting that the entire range of dorso-ventral identities can be generated within continuous neuroepithelia. Consistent with this, we demonstrate the presence of forebrain organizing centers that express secreted growth factors, which may be involved in dorso-ventral patterning within organoids. Furthermore, we demonstrate the timed generation of neurons with mature morphologies, as well as the subsequent generation of astrocytes and oligodendrocytes. Our work provides the methodology and quality criteria for phenotypic analysis of brain organoids and shows that the spatial and temporal patterning events governing human brain development can be recapitulated in vitro.
Bibliographical noteFunding Information:
We thank all members of the Knoblich, Lancaster, and Chung laboratories for discussions and support. We thank Raika Sieger for technical support. We are grateful to all IMP/IMBA and VBCF service facilities for providing technical support. We are particularly indebted to G. Petri and P. Pasierbek for imaging support and T. Lendl for help with 3D reconstructions. We thank Alex Phillips for generation of the membrane-targeted GFP construct. M.A.L. was funded by a Marie Curie Postdoctoral Fellowship. Work in M.A.L.'s laboratory is supported by the Medical Research Council MC_UP_1201/9. Work in J.A.K.'s laboratory is supported by the Austrian Academy of Sciences, the Austrian Science Fund (grants I_1281-B19 and Z_153_B09), and an advanced grant from the European Research Council (ERC). K.C. is supported by Burroughs Wellcome Fund Career Awards at the Scientific Interface, the Searle Scholars Program, Packard award in Science and Engineering, NARSAD Young Investigator Award, JPB Foundation (PIIF and PNDRF), NCSOFT Cultural Foundation, and NIH (1-U01-NS090473-01). Resources that may help enable general users to establish the SWITCH methodology are freely available online (http://www.chunglabresources.com).
© 2017 The Authors
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Immunology and Microbiology(all)