Generation and long-term culture of advanced cerebral organoids for studying later stages of neural development

Giandomenico, Stefano L.; Sutcliffe, Magdalena; Lancaster, Madeline A.

doi:10.1038/s41596-020-00433-w

Cited by 153 publications

(122 citation statements)

References 28 publications

Supporting

Mentioning

109

Contrasting

Order By: Relevance

“…Staining for both caspase 3/7 and TUNEL was found to be substantially lower/undetectable in the EBs cultured on the porous membrane when compared to those maintained as spheroids in suspension. These data indicate an improved cell viability, correlating with previous studies which undertook similar procedures to improve the local cellular microenvironment ( Giandomenico et al, 2019 , 2021 ).…”

Section: Discussionsupporting

confidence: 87%

“…To improve EB viability and allow for long term cell differentiation and maturation, EBs were seeded onto a porous polystyrene scaffold in order to facilitate a shape transformation and create a microenvironment with a more uniform distribution of nutrients and oxygen. Similar strategies are used in ex vivo organotypic slice culture and to extend the differentiation, complexity, and longevity of neurospheres ( Nestor et al, 2013 ; Giandomenico et al, 2019 , 2021 ); indeed, the use of complex 3D culture strategies combining biomaterial scaffolds with PSCs in order to direct and enhance their differentiation has expanded significantly over the last decade ( Willerth and Sakiyama-Elbert, 2019 ). Initial studies were carried out using a well characterised EC cell line ( Przyborski, 2001 ; Clarke et al, 2017 ), due to shared characteristics with ES cell populations ( Andrews et al, 2005 ) but simpler culture requirements, allowing for large numbers of EBs to be generated during method optimisation.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Using Advanced Cell Culture Techniques to Differentiate Pluripotent Stem Cells and Recreate Tissue Structures Representative of Teratoma Xenografts

Smith

Aguilar

Owens

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Various methods are currently used to investigate human tissue differentiation, including human embryo culture and studies utilising pluripotent stem cells (PSCs) such as in vitro embryoid body formation and in vivo teratoma assays. Each method has its own distinct advantages, yet many are limited due to being unable to achieve the complexity and maturity of tissue structures observed in the developed human. The teratoma xenograft assay allows maturation of more complex tissue derivatives, but this method has ethical issues surrounding animal usage and significant protocol variation. In this study, we have combined three-dimensional (3D) in vitro cell technologies including the common technique of embryoid body (EB) formation with a novel porous scaffold membrane, in order to prolong cell viability and extend the differentiation of PSC derived EBs. This approach enables the formation of more complex morphologically identifiable 3D tissue structures representative of all three primary germ layers. Preliminary in vitro work with the human embryonal carcinoma line TERA2.SP12 demonstrated improved EB viability and enhanced tissue structure formation, comparable to teratocarcinoma xenografts derived in vivo from the same cell line. This is thought to be due to reduced diffusion distances as the shape of the spherical EB transforms and flattens, allowing for improved nutritional/oxygen support to the developing structures over extended periods. Further work with EBs derived from murine embryonic stem cells demonstrated that the formation of a wide range of complex, recognisable tissue structures could be achieved within 2–3 weeks of culture. Rudimentary tissue structures from all three germ layers were present, including epidermal, cartilage and epithelial tissues, again, strongly resembling tissue structure of teratoma xenografts of the same cell line. Proof of concept work with EBs derived from the human embryonic stem cell line H9 also showed the ability to form complex tissue structures within this system. This novel yet simple model offers a controllable, reproducible method to achieve complex tissue formation in vitro. It has the potential to be used to study human developmental processes, as well as offering an animal free alternative method to the teratoma assay to assess the developmental potential of novel stem cell lines.

show abstract

Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Using Advanced Cell Culture Techniques to Differentiate Pluripotent Stem Cells and Recreate Tissue Structures Representative of Teratoma Xenografts

Smith

Aguilar

Owens

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

show abstract

“…To resolve the problem of central core necrosis of brain organoids, they are transplanted into the mouse brain as vascularized model of brain organoids in vivo (Mansour et al, 2018 ). In vitro organotypic slice culture improved long-term survival (Giandomenico et al, 2021 ), and air–liquid interface culture for cerebral organoids improved survival, axon outgrowth, and decreased apoptosis rate (Giandomenico et al, 2019 ). It might be the reason why VEGFA and HIF1A did not significantly change, and the lack of vascular system was one of the limitation of cerebral organoids.…”

Section: Discussionmentioning

confidence: 99%

Gene Expression Profiles of Human Cerebral Organoids Identify PPAR Pathway and PKM2 as Key Markers for Oxygen-Glucose Deprivation and Reoxygenation

Iwasa

Matsui

Iguchi

et al. 2021

Front. Cell. Neurosci.

View full text Add to dashboard Cite

Ischemic stroke is one of the most common neurological diseases. However, the impact of ischemic stroke on human cerebral tissue remains largely unknown due to a lack of ischemic human brain samples. In this study, we applied cerebral organoids derived from human induced pluripotent stem cells to evaluate the effect of oxygen-glucose deprivation/reoxygenation (OGD/R). Pathway analysis showed the relationships between vitamin digestion and absorption, fat digestion and absorption, peroxisome proliferator-activated receptor (PPAR) signaling pathway, and complement and coagulation cascades. Combinational verification with transcriptome and gene expression analysis of different cell types revealed fatty acids-related PPAR signaling pathway and pyruvate kinase isoform M2 (PKM2) as key markers of neuronal cells in response to OGD/R. These findings suggest that, although there remain some limitations to be improved, our ischemic stroke model using human cerebral organoids would be a potentially useful tool when combined with other conventional two-dimensional (2D) mono-culture systems.

show abstract

“…The majority of BOs generated using current protocols resemble an early stage of fetal brain [ 19 , 20 , 25 , 26 , 139 ], and which are not suitable to investigate the physiological and pathological aspects of adult brain, like the late-onset neurodegenerative diseases. The major reason is the BOs lack of a circulation system with blood vessels to continually supply of oxygen and nutrients for extended in vitro culture.…”

Section: Challenges For Existing Bo Modelsmentioning

confidence: 99%

Human pluripotent stem cell-derived brain organoids as in vitro models for studying neural disorders and cancer

Luo

2021

Cell Biosci

View full text Add to dashboard Cite

The sheer complexities of brain and resource limitation of human brain tissue greatly hamper our understanding of the brain disorders and cancers. Recently developed three-dimensional (3D) brain organoids (BOs) are self-organized and spontaneously differentiated from human pluripotent stem cells (hPSCs) in vitro, which exhibit similar features with cell type diversity, structural organization, and functional connectivity as the developing human brain. Based on these characteristics, hPSC-derived BOs (hPDBOs) provide new opportunities to recapitulate the complicated processes during brain development, neurodegenerative disorders, and brain cancers in vitro. In this review, we will provide an overview of existing BO models and summarize the applications of this technology in modeling the neural disorders and cancers. Furthermore, we will discuss the challenges associated with their use as in vitro models for disease modeling and the potential future direction.

show abstract

Generation and long-term culture of advanced cerebral organoids for studying later stages of neural development

Cited by 153 publications

References 28 publications

Using Advanced Cell Culture Techniques to Differentiate Pluripotent Stem Cells and Recreate Tissue Structures Representative of Teratoma Xenografts

Using Advanced Cell Culture Techniques to Differentiate Pluripotent Stem Cells and Recreate Tissue Structures Representative of Teratoma Xenografts

Gene Expression Profiles of Human Cerebral Organoids Identify PPAR Pathway and PKM2 as Key Markers for Oxygen-Glucose Deprivation and Reoxygenation

Human pluripotent stem cell-derived brain organoids as in vitro models for studying neural disorders and cancer

Contact Info

Product

Resources

About