Human stem cell based corneal tissue mimicking structures using laser-assisted 3D bioprinting and functional bioinks

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dc.identifier.uri Sorkio, Anni Koch, Lothar Koivusalo, Laura Deiwick, Andrea Miettinen, Susanna Chichkov, Boris Skottman, Heli 2018-06-27T11:45:28Z 2018-06-27T11:45:28Z 2018
dc.identifier.citation Sorkio, A.; Koch, L.; Koivusalo, L.; Deiwick, A.; Miettinen, S. et al.: Human stem cell based corneal tissue mimicking structures using laser-assisted 3D bioprinting and functional bioinks. In: Biomaterials 171 (2018), S. 57-71. DOI:
dc.description.abstract There is a high demand for developing methods to produce more native-like 3D corneal structures. In the present study, we produced 3D cornea-mimicking tissues using human stem cells and laser-assisted bioprinting (LaBP). Human embryonic stem cell derived limbal epithelial stem cells (hESC-LESC) were used as a cell source for printing epithelium-mimicking structures, whereas human adipose tissue derived stem cells (hASCs) were used for constructing layered stroma-mimicking structures. The development and optimization of functional bioinks was a crucial step towards successful bioprinting of 3D corneal structures. Recombinant human laminin and human sourced collagen I served as the bases for the functional bioinks. We used two previously established LaBP setups based on laser induced forward transfer, with different laser wavelengths and appropriate absorption layers. We bioprinted three types of corneal structures: stratified corneal epithelium using hESC-LESCs, lamellar corneal stroma using alternating acellular layers of bioink and layers with hASCs, and finally structures with both a stromal and epithelial part. The printed constructs were evaluated for their microstructure, cell viability and proliferation, and key protein expression (Ki67, p63α, p40, CK3, CK15, collagen type I, VWF). The 3D printed stromal constructs were also implanted into porcine corneal organ cultures. Both cell types maintained good viability after printing. Laser-printed hESC-LESCs showed epithelial cell morphology, expression of Ki67 proliferation marker and co-expression of corneal progenitor markers p63α and p40. Importantly, the printed hESC-LESCs formed a stratified epithelium with apical expression of CK3 and basal expression of the progenitor markers. The structure of the 3D bioprinted stroma demonstrated that the hASCs had organized horizontally as in the native corneal stroma and showed positive labeling for collagen I. After 7 days in porcine organ cultures, the 3D bioprinted stromal structures attached to the host tissue with signs of hASCs migration from the printed structure. This is the first study to demonstrate the feasibility of 3D LaBP for corneal applications using human stem cells and successful fabrication of layered 3D bioprinted tissues mimicking the structure of the native corneal tissue. eng
dc.language.iso eng
dc.publisher London : Elsevier Ltd.
dc.relation.ispartofseries Biomaterials 171 (2018)
dc.rights CC BY-NC-ND 4.0 Unported
dc.subject 3D bioprinting eng
dc.subject Adipose stem cells eng
dc.subject Cornea eng
dc.subject Human collagen I eng
dc.subject Human pluripotent stem cells eng
dc.subject Laser-assisted bioprinting eng
dc.subject Limbal epithelial stem cells eng
dc.subject Recombinant human laminin eng
dc.subject 3D printers eng
dc.subject Boron compounds eng
dc.subject Collagen eng
dc.subject Cytology eng
dc.subject Histology eng
dc.subject Lanthanum compounds eng
dc.subject Phosphorus compounds eng
dc.subject Tissue eng
dc.subject Adipose stem cells eng
dc.subject Bioprinting eng
dc.subject Cornea eng
dc.subject Human collagens eng
dc.subject Laminin eng
dc.subject Pluripotent stem cells eng
dc.subject Stem cells eng
dc.subject.ddc 570 | Biowissenschaften, Biologie ger
dc.title Human stem cell based corneal tissue mimicking structures using laser-assisted 3D bioprinting and functional bioinks
dc.type article
dc.type Text
dc.relation.issn 0142-9612
dc.bibliographicCitation.volume 171
dc.bibliographicCitation.firstPage 57
dc.bibliographicCitation.lastPage 71
dc.description.version publishedVersion
tib.accessRights frei zug�nglich

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