Silk scaffolding drives self-assembly of functional and mature human brain organoids
Human pluripotent stem cells (hPSCs) are intrinsically able to self-organize into cerebral organoids that mimic features of developing human brain tissue. These three-dimensional structures provide a unique opportunity to generate cytoarchitecture and cell-cell interactions reminiscent of human brai...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2022.1023279/full |
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| author | Edoardo Sozzi Janko Kajtez Andreas Bruzelius Milan Finn Wesseler Fredrik Nilsson Marcella Birtele Niels B. Larsen Daniella Rylander Ottosson Petter Storm Malin Parmar Alessandro Fiorenzano |
| author_facet | Edoardo Sozzi Janko Kajtez Andreas Bruzelius Milan Finn Wesseler Fredrik Nilsson Marcella Birtele Niels B. Larsen Daniella Rylander Ottosson Petter Storm Malin Parmar Alessandro Fiorenzano |
| author_sort | Edoardo Sozzi |
| collection | DOAJ |
| description | Human pluripotent stem cells (hPSCs) are intrinsically able to self-organize into cerebral organoids that mimic features of developing human brain tissue. These three-dimensional structures provide a unique opportunity to generate cytoarchitecture and cell-cell interactions reminiscent of human brain complexity in a dish. However, current in vitro brain organoid methodologies often result in intra-organoid variability, limiting their use in recapitulating later developmental stages as well as in disease modeling and drug discovery. In addition, cell stress and hypoxia resulting from long-term culture lead to incomplete maturation and cell death within the inner core. Here, we used a recombinant silk microfiber network as a scaffold to drive hPSCs to self-arrange into engineered cerebral organoids. Silk scaffolding promoted neuroectoderm formation and reduced heterogeneity of cellular organization within individual organoids. Bulk and single cell transcriptomics confirmed that silk cerebral organoids display more homogeneous and functionally mature neuronal properties than organoids grown in the absence of silk scaffold. Furthermore, oxygen sensing analysis showed that silk scaffolds create more favorable growth and differentiation conditions by facilitating the delivery of oxygen and nutrients. The silk scaffolding strategy appears to reduce intra-organoid variability and enhances self-organization into functionally mature human brain organoids. |
| first_indexed | 2024-04-12T13:36:54Z |
| format | Article |
| id | doaj.art-dc93822d588747b9a5b8c9265e0e8c2f |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-12T13:36:54Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-dc93822d588747b9a5b8c9265e0e8c2f2022-12-22T03:30:59ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2022-10-011010.3389/fcell.2022.10232791023279Silk scaffolding drives self-assembly of functional and mature human brain organoidsEdoardo Sozzi0Janko Kajtez1Andreas Bruzelius2Milan Finn Wesseler3Fredrik Nilsson4Marcella Birtele5Niels B. Larsen6Daniella Rylander Ottosson7Petter Storm8Malin Parmar9Alessandro Fiorenzano10Department of Experimental Medical Science, Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Experimental Medical Science, Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Experimental Medical Science, Regenerative Neurophysiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Health Technology (DTU Health Tech), Technical University of Denmark, Kongens Lyngby, DenmarkDepartment of Experimental Medical Science, Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Experimental Medical Science, Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Health Technology (DTU Health Tech), Technical University of Denmark, Kongens Lyngby, DenmarkDepartment of Experimental Medical Science, Regenerative Neurophysiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Experimental Medical Science, Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Experimental Medical Science, Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenDepartment of Experimental Medical Science, Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Lund University, Lund, SwedenHuman pluripotent stem cells (hPSCs) are intrinsically able to self-organize into cerebral organoids that mimic features of developing human brain tissue. These three-dimensional structures provide a unique opportunity to generate cytoarchitecture and cell-cell interactions reminiscent of human brain complexity in a dish. However, current in vitro brain organoid methodologies often result in intra-organoid variability, limiting their use in recapitulating later developmental stages as well as in disease modeling and drug discovery. In addition, cell stress and hypoxia resulting from long-term culture lead to incomplete maturation and cell death within the inner core. Here, we used a recombinant silk microfiber network as a scaffold to drive hPSCs to self-arrange into engineered cerebral organoids. Silk scaffolding promoted neuroectoderm formation and reduced heterogeneity of cellular organization within individual organoids. Bulk and single cell transcriptomics confirmed that silk cerebral organoids display more homogeneous and functionally mature neuronal properties than organoids grown in the absence of silk scaffold. Furthermore, oxygen sensing analysis showed that silk scaffolds create more favorable growth and differentiation conditions by facilitating the delivery of oxygen and nutrients. The silk scaffolding strategy appears to reduce intra-organoid variability and enhances self-organization into functionally mature human brain organoids.https://www.frontiersin.org/articles/10.3389/fcell.2022.1023279/fullhuman pluripotent stem cellscerebral organoidsilk scaffoldingtissue engineeringoxygen sensing |
| spellingShingle | Edoardo Sozzi Janko Kajtez Andreas Bruzelius Milan Finn Wesseler Fredrik Nilsson Marcella Birtele Niels B. Larsen Daniella Rylander Ottosson Petter Storm Malin Parmar Alessandro Fiorenzano Silk scaffolding drives self-assembly of functional and mature human brain organoids Frontiers in Cell and Developmental Biology human pluripotent stem cells cerebral organoid silk scaffolding tissue engineering oxygen sensing |
| title | Silk scaffolding drives self-assembly of functional and mature human brain organoids |
| title_full | Silk scaffolding drives self-assembly of functional and mature human brain organoids |
| title_fullStr | Silk scaffolding drives self-assembly of functional and mature human brain organoids |
| title_full_unstemmed | Silk scaffolding drives self-assembly of functional and mature human brain organoids |
| title_short | Silk scaffolding drives self-assembly of functional and mature human brain organoids |
| title_sort | silk scaffolding drives self assembly of functional and mature human brain organoids |
| topic | human pluripotent stem cells cerebral organoid silk scaffolding tissue engineering oxygen sensing |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2022.1023279/full |
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