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1.	Buchmann, Sebastian, et al. (author) Probabilistic cell seeding and non-autofluorescent 3D-printed structures as scalable approach for multi-level co-culture modeling 2023 In: Materials Today Bio. - : Elsevier BV. - 2590-0064. ; 21, s. 100706-100706 Journal article (peer-reviewed)abstract To model complex biological tissue in vitro, a specific layout for the position and numbers of each cell type isnecessary. Establishing such a layout requires manual cell placement in three dimensions (3D) with micrometricprecision, which is complicated and time-consuming. Moreover, 3D printed materials used in compartmentalizedmicrofluidic models are opaque or autofluorescent, hindering parallel optical readout and forcing serial charac-terization methods, such as patch-clamp probing. To address these limitations, we introduce a multi-level co-culture model realized using a parallel cell seeding strategy of human neurons and astrocytes on 3D structuresprinted with a commercially available non-autofluorescent resin at micrometer resolution. Using a two-stepstrategy based on probabilistic cell seeding, we demonstrate a human neuronal monoculture that forms net-works on the 3D printed structure and can establish cell-projection contacts with an astrocytic-neuronal co-cultureseeded on the glass substrate. The transparent and non-autofluorescent printed platform allows fluorescence-based immunocytochemistry and calcium imaging. This approach provides facile multi-level compartmentaliza-tion of different cell types and routes for pre-designed cell projection contacts, instrumental in studying complextissue, such as the human brain.

Buchmann, Sebastian, et al. (author)
Probabilistic cell seeding and non-autofluorescent 3D-printed structures as scalable approach for multi-level co-culture modeling
2023
In: Materials Today Bio. - : Elsevier BV. - 2590-0064. ; 21, s. 100706-100706
Journal article (peer-reviewed)abstract
- To model complex biological tissue in vitro, a specific layout for the position and numbers of each cell type isnecessary. Establishing such a layout requires manual cell placement in three dimensions (3D) with micrometricprecision, which is complicated and time-consuming. Moreover, 3D printed materials used in compartmentalizedmicrofluidic models are opaque or autofluorescent, hindering parallel optical readout and forcing serial charac-terization methods, such as patch-clamp probing. To address these limitations, we introduce a multi-level co-culture model realized using a parallel cell seeding strategy of human neurons and astrocytes on 3D structuresprinted with a commercially available non-autofluorescent resin at micrometer resolution. Using a two-stepstrategy based on probabilistic cell seeding, we demonstrate a human neuronal monoculture that forms net-works on the 3D printed structure and can establish cell-projection contacts with an astrocytic-neuronal co-cultureseeded on the glass substrate. The transparent and non-autofluorescent printed platform allows fluorescence-based immunocytochemistry and calcium imaging. This approach provides facile multi-level compartmentaliza-tion of different cell types and routes for pre-designed cell projection contacts, instrumental in studying complextissue, such as the human brain.

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