3D Bioprinting of Murine Cortical Astrocytes for Engineering Neural-Like Tissue

Cruz, Elisa M.; Ribeiro, Tais Novaki; Mundim, Mayara V; Porcionatto, Marimélia

doi:10.3791/62691

Cited by 7 publications

(6 citation statements)

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“…Despite concerns surrounding reductions in cell viability due to shear stressors during bioprinting (Shi et al, 2018), we were able to successfully bioprint iPSC-derived astrocytes, NPCs and iBMECs within our lead matrix with a good level of viability similar to what we found using manual gelation. Recent work using extrusion-based bioprinting of rodent astrocytes within composite gelatin-methacryloyl matrices and NPCs within collagen-based matrices have reported viability ranging from 75 - 90 %, similar to our results (de Melo, Cruz, Ribeiro, Mundim, & Porcionatto, 2021; Ouyang et al, 2020; Salaris et al, 2019; Sharma, Smits, De La Vega, Lee, & Willerth, 2020). Furthermore, we show that iBMECs bioprinted in our lead matrix, cultured in AM with tapered ROCK inhibition, remain viable up to 4 weeks.…”

Section: Discussionsupporting

confidence: 91%

3D Bioprinting of Stem Cell-Derived Central Nervous System Cells Enables Astrocyte Growth, Vasculogenesis and Enhances Neural Differentiation/Function

Sullivan

Lane

Volkerling³

et al. 2022

Preprint

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Current research tools for pre-clinical drug development such as rodent models and 2D immortalised monocultures have failed to serve as effective translational models for human CNS disorders. Recent advancements in the development of iPSCs and 3D culturing can improve thein vivo-relevanceof pre-clinical models, while generating 3D cultures though novel bioprinting technologies can offer increased scalability and replicability. As such, there is a need to develop platforms that combine iPSC-derived cells with 3D bioprinting to produce scalable, tunable and biomimetic cultures for preclinical drug discovery applications. We report a biocompatible PEG-based matrix which incorporates RGD and YIGSR peptide motifs and full length collagen IV at a stiffness similar to the human brain (1.5 kPa). Using a high-throughput commercial bioprinter we report the viable culture and morphological development of iPSC-derived astrocytes, brain microvascular endothelial cells, neural progenitors and neurons in our novel matrix. We also show that this system supports endothelial vasculogenesis and enhances neural differentiation and spontaneous activity. This platform forms a foundation for more complex, multicellular models to facilitate high-throughput translational drug discovery for CNS disorders.

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Section: Discussionsupporting

confidence: 91%

3D Bioprinting of Stem Cell-Derived Central Nervous System Cells Enables Astrocyte Growth, Vasculogenesis and Enhances Neural Differentiation/Function

Sullivan

Lane

Volkerling³

et al. 2022

Preprint

View full text Add to dashboard Cite

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“…Despite concerns surrounding reductions in cell viability due to shear stressors during bioprinting (Shi et al, 2018), we were able to successfully bioprint iPSC-derived astrocytes, NPCs and iBMECs within our lead matrix with a good level of viability similar to what we found using manual gelation. Recent work using extrusion-based bioprinting of rodent astrocytes within composite gelatin-methacryloyl matrices and NPCs within collagenbased matrices have reported viability ranging from 75 -90 %, similar to our results (de Melo, Cruz, Ribeiro, Mundim, & Porcionatto, 2021;Ouyang et al, 2020;Salaris et al, 2019;Sharma, Smits, De La Vega, Lee, & Willerth, 2020). Furthermore, we show that iBMECs bioprinted in our lead matrix, cultured in AM with tapered ROCK inhibition, remain viable up to 4 weeks.…”

Section: Discussionsupporting

confidence: 91%

3D Bioprinting of Stem Cell-Derived Central Nervous System Cells Enables Astrocyte Growth, Vasculogenesis and Enhances Neural Differentiation/Function

Sullivan

Lane

Volkerling

et al. 2022

Preprint

View full text Add to dashboard Cite

Current research tools for pre-clinical drug development such as rodent models and 2D immortalised monocultures have failed to serve as effective translational models for human CNS disorders. Recent advancements in the development of iPSCs and 3D culturing can improve the in vivo-relevance of pre-clinical models, while generating 3D cultures though novel bioprinting technologies can offer increased scalability and replicability. As such, there is a need to develop platforms that combine iPSC-derived cells with 3D bioprinting to produce scalable, tunable and biomimetic cultures for preclinical drug discovery applications. We report a biocompatible PEG-based matrix which incorporates RGD and YIGSR peptide motifs and full length collagen IV at a stiffness similar to the human brain (1.5 kPa). Using a high-throughput commercial bioprinter we report the viable culture and morphological development of iPSC-derived astrocytes, brain microvascular endothelial cells, neural progenitors and neurons in our novel matrix. We also show that this system supports endothelial vasculogenesis and enhances neural differentiation and spontaneous activity. This platform forms a foundation for more complex, multicellular models to facilitate high-throughput translational drug discovery for CNS disorders.

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Section: Resultsmentioning

confidence: 99%

“…Bioinks Preparation and Crosslinking: GelMA was synthesized according to the recently published protocol. [45] Two bioinks were prepared: GGFL, composed of gelatin (Sigma-Aldrich) 4% (m/v), GelMA 2% (m/v), fibrinogen (Sigma-Aldrich) 3 mg mL -1 , and laminin 2 µg mL -1 , and GGF, composed of gelatin 5% (m/v), GelMA 5% (m/v), and fibrinogen 1.5 mg mL -1 . Briefly, gelatin and GelMA were diluted in PBS 1×, and fibrinogen previously prepared in saline solution was mixed to the gelatin-GelMA emulsion until the established concentrations.…”

Section: Methodsmentioning

confidence: 99%

“…Primary culture of astrocytes derived from C57Bl/6 mice is well established in the literature and has been a target cell type in regenerative neurobiology studies, including the biofabrication of 3D in vitro neural models. [45,46] Initially, cells were mixed to the biomaterials' solution, bioprinted, and cultured at 37 °C and 5% of CO 2 (Figure 2A). The bioprinted structure possessed a squared shape, and cells were homogeneously distributed within the bioprinted layers (Figure 2B).…”

Section: Characterization Of Bioinks' Biological Propertiesmentioning

confidence: 99%

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3D Bioprinted Neural‐Like Tissue as a Platform to Study Neurotropism of Mouse‐Adapted SARS‐CoV‐2

et al. 2022

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The effects of neuroinvasion by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) become clinically relevant due to the numerous neurological symptoms observed in Corona Virus Disease 2019 (COVID‐19) patients during infection and post‐COVID syndrome or long COVID. This study reports the biofabrication of a 3D bioprinted neural‐like tissue as a proof‐of‐concept platform for a more representative study of SARS‐CoV‐2 brain infection. Bioink is optimized regarding its biophysical properties and is mixed with murine neural cells to construct a 3D model of COVID‐19 infection. Aiming to increase the specificity to murine cells, SARS‐CoV‐2 is mouse‐adapted (MA‐SARS‐CoV‐2) in vitro, in a protocol first reported here. MA‐SARS‐CoV‐2 reveals mutations located at the Orf1a and Orf3a domains and is evolutionarily closer to the original Wuhan SARS‐CoV‐2 strain than SARS‐CoV‐2 used for adaptation. Remarkably, MA‐SARS‐CoV‐2 shows high specificity to murine cells, which present distinct responses when cultured in 2D and 3D systems, regarding cell morphology, neuroinflammation, and virus titration. MA‐SARS‐CoV‐2 represents a valuable tool in studies using animal models, and the 3D neural‐like tissue serves as a powerful in vitro platform for modeling brain infection, contributing to the development of antivirals and new treatments for COVID‐19.

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3D Bioprinting of Murine Cortical Astrocytes for Engineering Neural-Like Tissue

Cited by 7 publications

References 0 publications

3D Bioprinting of Stem Cell-Derived Central Nervous System Cells Enables Astrocyte Growth, Vasculogenesis and Enhances Neural Differentiation/Function

3D Bioprinting of Stem Cell-Derived Central Nervous System Cells Enables Astrocyte Growth, Vasculogenesis and Enhances Neural Differentiation/Function

3D Bioprinting of Stem Cell-Derived Central Nervous System Cells Enables Astrocyte Growth, Vasculogenesis and Enhances Neural Differentiation/Function

3D Bioprinted Neural‐Like Tissue as a Platform to Study Neurotropism of Mouse‐Adapted SARS‐CoV‐2

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