Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body Array
Stem cell technology and embryonic stem cell models are of great interest in biomedical research since they provide deeper insights into, e.g., neurogenesis and early mammalian brain development. Despite their great scientific potential, the reliable establishment of three-dimensional embryoid bodie...
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MDPI AG
2022-01-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/14/2/339 |
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| author | Christoph Eilenberger Mario Rothbauer Konstanze Brandauer Sarah Spitz Eva-Kathrin Ehmoser Seta Küpcü Peter Ertl |
| author_facet | Christoph Eilenberger Mario Rothbauer Konstanze Brandauer Sarah Spitz Eva-Kathrin Ehmoser Seta Küpcü Peter Ertl |
| author_sort | Christoph Eilenberger |
| collection | DOAJ |
| description | Stem cell technology and embryonic stem cell models are of great interest in biomedical research since they provide deeper insights into, e.g., neurogenesis and early mammalian brain development. Despite their great scientific potential, the reliable establishment of three-dimensional embryoid bodies (EBs) remains a major challenge, and the current lack of standardization and comparability is still limiting a broader application and translation of stem cell technology. Among others, a vital aspect for the reliable formation of EBs is optimizing differentiation protocols since organized differentiation is influenced by soluble inducers and EB size. A microfluidic biochip array was employed to automate cell loading and optimize directed neuronal and astrocytic differentiation protocols using murine P19 embryoid bodies to facilitate reliable embryonic stem cell differentiation. Our gravity-driven microfluidic size-controlled embryoid body-on-a-chip system allows (a) the robust operation and cultivation of up to 90 EBs in parallel and (b) the reproducible generation of five increasing sizes ranging from 300 µm to 1000 µm diameters. A comparative study adds two differentiation-inducers such as retinoic acid and EC23 to size-controlled embryoid bodies to identify the optimal differentiation protocol. Our study revealed a 1.4 to 1.9-fold higher neuron and astrocyte expression in larger embryoid bodies (above 750 µm) over smaller-sized EBs (below 450 µm), thus highlighting the importance of EB size in the establishment of robust neurodevelopmental in vitro models. |
| first_indexed | 2024-03-09T21:14:46Z |
| format | Article |
| id | doaj.art-3088e6ad07624e448c35ac9a33926f07 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T21:14:46Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-3088e6ad07624e448c35ac9a33926f072023-11-23T21:37:40ZengMDPI AGPharmaceutics1999-49232022-01-0114233910.3390/pharmaceutics14020339Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body ArrayChristoph Eilenberger0Mario Rothbauer1Konstanze Brandauer2Sarah Spitz3Eva-Kathrin Ehmoser4Seta Küpcü5Peter Ertl6Faculty of Technical Chemistry, Vienna University of Technology, Getreidemarkt 9, 1060 Vienna, AustriaFaculty of Technical Chemistry, Vienna University of Technology, Getreidemarkt 9, 1060 Vienna, AustriaFaculty of Technical Chemistry, Vienna University of Technology, Getreidemarkt 9, 1060 Vienna, AustriaFaculty of Technical Chemistry, Vienna University of Technology, Getreidemarkt 9, 1060 Vienna, AustriaDepartment of Nanobiotechnology, Institute of Synthetic Bioarchitectures, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, AustriaDepartment of Nanobiotechnology, Institute of Synthetic Bioarchitectures, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, AustriaFaculty of Technical Chemistry, Vienna University of Technology, Getreidemarkt 9, 1060 Vienna, AustriaStem cell technology and embryonic stem cell models are of great interest in biomedical research since they provide deeper insights into, e.g., neurogenesis and early mammalian brain development. Despite their great scientific potential, the reliable establishment of three-dimensional embryoid bodies (EBs) remains a major challenge, and the current lack of standardization and comparability is still limiting a broader application and translation of stem cell technology. Among others, a vital aspect for the reliable formation of EBs is optimizing differentiation protocols since organized differentiation is influenced by soluble inducers and EB size. A microfluidic biochip array was employed to automate cell loading and optimize directed neuronal and astrocytic differentiation protocols using murine P19 embryoid bodies to facilitate reliable embryonic stem cell differentiation. Our gravity-driven microfluidic size-controlled embryoid body-on-a-chip system allows (a) the robust operation and cultivation of up to 90 EBs in parallel and (b) the reproducible generation of five increasing sizes ranging from 300 µm to 1000 µm diameters. A comparative study adds two differentiation-inducers such as retinoic acid and EC23 to size-controlled embryoid bodies to identify the optimal differentiation protocol. Our study revealed a 1.4 to 1.9-fold higher neuron and astrocyte expression in larger embryoid bodies (above 750 µm) over smaller-sized EBs (below 450 µm), thus highlighting the importance of EB size in the establishment of robust neurodevelopmental in vitro models.https://www.mdpi.com/1999-4923/14/2/339embryoid bodymicrofluidicsorgan-on-a-chipneural differentiationEC23retinoic acid |
| spellingShingle | Christoph Eilenberger Mario Rothbauer Konstanze Brandauer Sarah Spitz Eva-Kathrin Ehmoser Seta Küpcü Peter Ertl Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body Array Pharmaceutics embryoid body microfluidics organ-on-a-chip neural differentiation EC23 retinoic acid |
| title | Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body Array |
| title_full | Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body Array |
| title_fullStr | Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body Array |
| title_full_unstemmed | Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body Array |
| title_short | Screening for Best Neuronal-Glial Differentiation Protocols of Neuralizing Agents Using a Multi-Sized Microfluidic Embryoid Body Array |
| title_sort | screening for best neuronal glial differentiation protocols of neuralizing agents using a multi sized microfluidic embryoid body array |
| topic | embryoid body microfluidics organ-on-a-chip neural differentiation EC23 retinoic acid |
| url | https://www.mdpi.com/1999-4923/14/2/339 |
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