T5 Towards human lung regeneration in end-stage respiratory failure: genetically-modifiable 3d organoid culture of human embryonic lung stem cells enables for the first time the study of human lung development in vitro

AbstractCerebral organoids provide unparalleled access to human brain development in vitro. However, variability induced by current culture methodologies precludes using organoids as robust disease models. To address this, we developed an automated Organoid Culture and Assay (ORCA) system to support longitudinal unbiased phenotyping of organoids at scale across multiple patient lines. We then characterized organoid variability using novel machine learning methods and found that the contribution of donor, clone, and batch is significant and remarkably consistent over gene expression, morphology, and cell-type composition. Next, we performed multi-factorial protocol optimization, producing a directed forebrain protocol compatible with 96-well culture that exhibits low variability while preserving tissue complexity. Finally, we used ORCA to study tuberous sclerosis, a disease with known genetics but poorly representative animal models. For the first time, we report highly reproducible early morphological and molecular signatures of disease in heterozygous TSC+/− forebrain organoids, demonstrating the benefit of a scaled organoid system for phenotype discovery in human disease models.

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Influence of corticosteroids and thyroxine on lung development in vitro

Acta Endocrinologica ◽

10.1530/acta.0.109s035-a ◽

1985 ◽

Vol 110 (1_Suppla) ◽

pp. S35-S36 ◽

Cited By ~ 3

Author(s):

B. ZIMMERMANN

Keyword(s):

Lung Development ◽

Development In Vitro

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Human embryonic lung epithelial tips are multipotent progenitors that can be expanded in vitro as long-term self-renewing organoids

eLife ◽

10.7554/elife.26575 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 84

Author(s):

Marko Z Nikolić ◽

Oriol Caritg ◽

Quitz Jeng ◽

Jo-Anne Johnson ◽

Dawei Sun ◽

...

Keyword(s):

Lung Development ◽

Mouse Lung ◽

Lung Epithelium ◽

Human Embryonic Lung ◽

Self Renewal ◽

Embryonic Mouse ◽

Multipotent Progenitors ◽

Lung Epithelial

The embryonic mouse lung is a widely used substitute for human lung development. For example, attempts to differentiate human pluripotent stem cells to lung epithelium rely on passing through progenitor states that have only been described in mouse. The tip epithelium of the branching mouse lung is a multipotent progenitor pool that self-renews and produces differentiating descendants. We hypothesized that the human distal tip epithelium is an analogous progenitor population and tested this by examining morphology, gene expression and in vitro self-renewal and differentiation capacity of human tips. These experiments confirm that human and mouse tips are analogous and identify signalling pathways that are sufficient for long-term self-renewal of human tips as differentiation-competent organoids. Moreover, we identify mouse-human differences, including markers that define progenitor states and signalling requirements for long-term self-renewal. Our organoid system provides a genetically-tractable tool that will allow these human-specific features of lung development to be investigated.

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