Assembly of Human Stem Cell-Derived Cortical Spheroids and Vascular Spheroids to Model 3-D Brain-like Tissues

Song, Liqing; Yuan, Xuegang; Jones, Zachary; Griffin, Kyle; Zhou, Yi; Ma, Teng; Li, Yan

doi:10.1038/s41598-019-42439-9

Cited by 118 publications

(101 citation statements)

References 83 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Regarding the current state of hydrogel use for brain in vitro cell research, the most popular hydrogels, according to the number of publications, are the cell culture-derived matrices Geltrex TM and Matrigel ® . They are used to develop human brain organoids from iPSCs and even applied for neural tissue engineering in animal studies [50][51][52]. The hydrogels well-support (although not so fast) neural cell differentiation and functional maturation [53][54][55].…”

Section: Discussionmentioning

confidence: 99%

Cerebellar Cells Self-Assemble into Functional Organoids on Synthetic, Chemically Crosslinked ECM-Mimicking Peptide Hydrogels

et al. 2020

View full text Add to dashboard Cite

Hydrogel-supported neural cell cultures are more in vivo-relevant compared to monolayers formed on glass or plastic substrates. However, there is a lack of synthetic microenvironment available for obtaining standardized and easily reproducible cultures characterized by tissue-mimicking cell composition, cell–cell interactions, and functional networks. Synthetic peptides representing the biological properties of the extracellular matrix (ECM) proteins have been reported to promote the adhesion-driven differentiation and functional maturation of neural cells. Thus, such peptides can serve as building blocks for engineering a standardized, all-synthetic environment. In this study, we have compared the effect of two chemically crosslinked hydrogel compositions on primary cerebellar cells: collagen-like peptide (CLP), and CLP with an integrin-binding motif arginine-glycine-aspartate (CLP-RGD), both conjugated to polyethylene glycol molecular templates (PEG-CLP and PEG-CLP-RGD, respectively) and fabricated as self-supporting membranes. Both compositions promoted a spontaneous organization of primary cerebellar cells into tissue-like clusters with fast-rising Ca2+ signals in soma, reflecting action potential generation. Notably, neurons on PEG-CLP-RGD had more neurites and better synaptic efficiency compared to PEG-CLP. For comparison, poly-L-lysine-coated glass and plastic surfaces did not induce formation of such spontaneously active networks. Additionally, contrary to the hydrogel membranes, glass substrates functionalized with PEG-CLP and PEG-CLP-RGD did not sufficiently support cell attachment and, subsequently, did not promote functional cluster formation. These results indicate that not only chemical composition but also the hydrogel structure and viscoelasticity are essential for bioactive signaling. The synthetic strategy based on ECM-mimicking, multifunctional blocks in registry with chemical crosslinking for obtaining tissue-like mechanical properties is promising for the development of fast and well standardized functional in vitro neural models and new regenerative therapies.

show abstract

Section: Discussionmentioning

confidence: 99%

Cerebellar Cells Self-Assemble into Functional Organoids on Synthetic, Chemically Crosslinked ECM-Mimicking Peptide Hydrogels

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Importantly, it has become clear in the last decade that cells sense and respond to the dimensionality and rigidity of their environment, and these qualities cannot be modeled using regular tissue culture methods 31 . Multicellular spheroid systems consisting of human primary or iPSC-derived EC, pericytes, astrocytes and neurons are cultured into multicellular BBB-and/or brain-organoid structures [32][33][34][35][36] . These organoids can be maintained for extensive time in culture, holding great promise to study neuronal functions.…”

Section: Discussionmentioning

confidence: 99%

An in vitro bioengineered model of the human arterial neurovascular unit to study neurodegenerative diseases

Robert

Weilinger

Zao

et al. 2020

Preprint

View full text Add to dashboard Cite

Introduction: The neurovascular unit (NVU) – the interaction between the neurons and the cerebrovasculature – is increasingly important to interrogate through human-based experimental models. Although advanced models of cerebral capillaries have been developed in the last decade, there is currently noin vitro3-dimensional (3D) perfusible model of the human cortical arterial NVU. Method: We used a tissue-engineering technique to develop a scaffold-directed, perfusible, 3D human NVU that is cultured in native-like flow conditions that mimics the anatomy and physiology of cortical penetrating arteries. Results: This system, composed of primary human vascular cells (endothelial cells, smooth muscle cells and astrocytes) and induced pluripotent stem cell (iPSC) derived neurons, demonstrates a physiological multilayer organization of the involved cell types. It also reproduces key characteristics of cortical neurons and astrocytes, as well as the formation of a selective and functional endothelial barrier. We further provide proof-of-principle that our in vitro human arterial NVU may be suitable to study neurodegenerative diseases such as Alzheimer’s disease (AD), as we report both phosphorylated tau and beta-amyloid accumulation in our model over time. Finally, we show that our arterial NVU model enables the study of neuronal and glial fluid biomarkers. Conclusion: This model is a suitable tool to investigate arterial NVU functions such as neuronal electrophysiology in health and disease. Further the design of platform allows culture under native-like flow conditions for extended periods of time and yields sufficient tissue and media for downstream immunohistochemistry and biochemistry analyses.

show abstract

“…Importantly, it has become clear in the last decade that cells sense and respond to the dimensionality and rigidity of their environment, and these qualities cannot be modeled using classical tissue culture methods 34 . Newer approaches often use multicellular spheroid systems consisting of human primary or iPSC-derived EC, pericytes, astrocytes and neurons that are cultured into multicellular BBB-and/or brain-organoid structures [35][36][37][38][39] . These organoids can be maintained for extensive time in culture, holding great promise to study neuronal functions.…”

Section: Discussionmentioning

confidence: 99%

An in vitro bioengineered model of the human arterial neurovascular unit to study neurodegenerative diseases

Robert

Weilinger

Zao

et al. 2020

Preprint

View full text Add to dashboard Cite

Introduction: The neurovascular unit (NVU) – the interaction between the neurons and the cerebrovasculature – is increasingly important to interrogate through human-based experimental models. Although advanced models of cerebral capillaries have been developed in the last decade, there is currently no in vitro 3-dimensional (3D) perfusible model of the human cortical arterial NVU.Method: We used a tissue-engineering technique to develop a scaffold-directed, perfusible, 3D human NVU that is cultured in native-like flow conditions that mimics the anatomy and physiology of cortical penetrating arteries.Results: This system, composed of primary human vascular cells (endothelial cells, smooth muscle cells and astrocytes) and induced pluripotent stem cell (iPSC) derived neurons, demonstrates a physiological multilayer organization of the involved cell types. It reproduces key characteristics of cortical neurons and astrocytes and enables formation of a selective and functional endothelial barrier. We provide proof-of-principle data showing that this in vitro human arterial NVU may be suitable to study neurovascular components of neurodegenerative diseases such as Alzheimer’s disease (AD), as endogenously produced phosphorylated tau and beta-amyloid accumulate in the model over time. Finally, neuronal and glial fluid biomarkers relevant to neurodegenerative diseases are measurable in our arterial NVU model.Conclusion: This model is a suitable research tool to investigate arterial NVU functions in healthy and disease states. Further, the design of the platform allows culture under native-like flow conditions for extended periods of time and yields sufficient tissue and media for downstream immunohistochemistry and biochemistry analyses.

show abstract

Assembly of Human Stem Cell-Derived Cortical Spheroids and Vascular Spheroids to Model 3-D Brain-like Tissues

Cited by 118 publications

References 83 publications

Cerebellar Cells Self-Assemble into Functional Organoids on Synthetic, Chemically Crosslinked ECM-Mimicking Peptide Hydrogels

Cerebellar Cells Self-Assemble into Functional Organoids on Synthetic, Chemically Crosslinked ECM-Mimicking Peptide Hydrogels

An in vitro bioengineered model of the human arterial neurovascular unit to study neurodegenerative diseases

An in vitro bioengineered model of the human arterial neurovascular unit to study neurodegenerative diseases

Contact Info

Product

Resources

About