Automation of Organoid Cultures: Current Protocols and Applications

Louey, Alexandra; Hernández, Damián; Pébay, Alice; Daniszewski, Maciej

doi:10.1177/24725552211024547

Cited by 10 publications

(5 citation statements)

References 125 publications

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“…This is due to issues with safety, scalability and serum batch-to-batch heterogeneity as well as difficulty dissociating cells from 3D organoids. In addition, automation of 3D organoid generation to produce large-scale reproducible organoids is a nascent field and will require further optimization to minimize variability across differentiation batches 47 . The method proposed here circumvents many of these issues and increases its amenability for manufacturing cell therapies at scale.…”

Section: Discussionmentioning

confidence: 99%

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

Fong,

Mendel,

Jascur

et al. 2023

Preprint

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iPSCs can serve as a renewable source of a consistent edited cell product, overcoming limitations of primary cells. While feeder-free generation of clinical grade iPSC-derived CD8 T cells has been achieved, differentiation of iPSC-derived CD4sp and regulatory T cells requires mouse stromal cells in an artificial thymic organoid. Here we report a serum- and feeder-free differentiation process suitable for large-scale production. Using an optimized concentration of PMA/Ionomycin, we generated iPSC-CD4sp T cells at high efficiency and converted them to Tregs using TGFβ and ATRA. Using zinc finger nucleases, we demonstrated high non-viral, targeted integration of an HLA-A2 CAR in iPSCs. iPSC-Tregs +/- HLA-A2 CAR phenotypically, transcriptionally and functionally resemble primary Tregs and suppress T cell proliferation in vitro. Our work is the first to demonstrate an iPSC-based platform amenable to manufacturing CD4 T cells to complement iPSC-CD8 oncology products and functional iPSC-Tregs to deliver Treg cell therapies at scale.

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Section: Discussionmentioning

confidence: 99%

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

Fong,

Mendel,

Jascur

et al. 2023

Preprint

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“…Up now, several automated culture systems have been developed, such as cerebral and retinal organoids. [ 62 ] Taking cerebral organoid derived from hiPSCs as an example, large neuroepithelial buds are observed from the EBs in automated culture, indicating the permissive conditions for cerebral organoid generation and maintenance. Image analysis tools can efficiently measure organoid diameters over time, and enable the train of the model by excluding cell debris, dust particles and imaging artifacts.…”

Section: Engineering Organoid Formationmentioning

confidence: 99%

Engineered Human Organoids for Biomedical Applications

Zhu,

Sun,

et al. 2023

Adv Funct Materials

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Human organoid models potentially offer a physiologically relevant platform to replace traditional monolayer cultures and animal models. In particular, the rapid development of engineered strategies including microfluidics, hydrogel, 3D printing and others, which have enormous advantages in comparison to conventional methods, is expected to further advance organoid technology. Up to now, many studies have demonstrated the engineered organoid models with complex cell composition, controlled structure, enhanced maturation, reduced heterogeneity, and so on. These engineered organoids are high promising for studies in development, disease, tissue repair, precision medicine and drug screening. In this review, a comprehensive summary of the engineered organoid model systems is provided based on microfluidics, hydrogel, 3D printing and so on. Then, the biomedical applications of these models are highlighted, which have displayed the great power in organoid field. Finally, the key bottlenecks and future development of organoid models are discussed about from an engineering perspective.

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“…Considering the limitations in conventional models, a human pluripotent stem cell-derived organoid is a promising new in vitro technique that could address these gaps. Being an in vitro model, the 3D organoid technology is more scalable and cost-effective compared to animal models [ 11 , 29 , 30 ]. Other than the practical considerations, the properties of a 3D organoid supersede its 2D counterpart in terms of physiological relevance.…”

Section: Comparison Between Models Of Infectious Diseasesmentioning

confidence: 99%

3D Human Organoids: The Next “Viral” Model for the Molecular Basis of Infectious Diseases

2022

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The COVID-19 pandemic has driven the scientific community to adopt an efficient and reliable model that could keep up with the infectious disease arms race. Coinciding with the pandemic, three dimensional (3D) human organoids technology has also gained traction in the field of infectious disease. An in vitro construct that can closely resemble the in vivo organ, organoid technology could bridge the gap between the traditional two-dimensional (2D) cell culture and animal models. By harnessing the multi-lineage characteristic of the organoid that allows for the recapitulation of the organotypic structure and functions, 3D human organoids have emerged as an essential tool in the field of infectious disease research. In this review, we will be providing a comparison between conventional systems and organoid models. We will also be highlighting how organoids played a role in modelling common infectious diseases and molecular mechanisms behind the pathogenesis of causative agents. Additionally, we present the limitations associated with the current organoid models and innovative strategies that could resolve these shortcomings.

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Automation of Organoid Cultures: Current Protocols and Applications

Cited by 10 publications

References 125 publications

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

Engineered Human Organoids for Biomedical Applications

3D Human Organoids: The Next “Viral” Model for the Molecular Basis of Infectious Diseases

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