Single cell transcriptomics captures features of developing and mature DA  neurons in human brain organoids and reveals more precise and reproducible patterning in silk-bioengineered culture

ABSTRACT: The inaccessibility of human tissue and the difficulty to achieve neuronal maturation and function in 2D cultures makes the study of human brain development, function and disease challenging. Here, we differentiated human pluripotent stem cells into three-dimensional (3D) regionalized human brain organoids which, when patterned towards a ventral midbrain (VM) fate, gave rise to mature and functional pigmented dopamine (DA) neurons. Using single cell transcriptomics, we defined the cellular composition and reconstructed the developmental trajectory of the different cell types, including DA neurons, by analyzing 120,000 cells harvested at different time points during differentiation. We also obtained an unbiased and comprehensive characterization of human DA subgroups, which reveals a molecular signature of DA neuron diversity and provides a valuable asset for the design of more targeted and effective stem-cell based therapies in Parkinson´s disease. However, the value of brain organoids in modeling later developmental stages and more mature neurons in a dish is hampered by their poor reproducibility and incomplete maturation. To address this issue, we designed a novel technological approach to generate bioengineered VM organoids supported by recombinant spider silk microfibers functionalized with full-length human laminin. Silk-VM organoids reproduce key molecular aspects of DA neurogenesis and, unlike conventional 3D culture, sustain the generation of functional DA neurons homogeneously throughout the organoid. Silk-VM organoids represent a significant advancement in the field of 3D culture, by better recapitulating physiologically relevant aspects of developing human brain tissue and thus paving the way toward their use in developmental studies as well as in in vitro disease modeling and drug discovery.

ORGANISM(S): Homo sapiens

PROVIDER: GSE168323 | GEO | 2021/10/29

REPOSITORIES: GEO