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Standardized 3D Bioprinting of Soft Tissue Models with Human Primary Cells

Rimann M1, Bono E2, Annaheim H2, Bleisch M2, Graf-Hausner U2.

J Lab Autom. 2015 Jan 21. pii: 2211068214567146.

1Institute of Chemistry and Biological Chemistry (ICBC), Zurich University of Applied Sciences (ZHAW), Waedenswil, Switzerland [email protected]

2Institute of Chemistry and Biological Chemistry (ICBC), Zurich University of Applied Sciences (ZHAW), Waedenswil, Switzerland.

Abstract

Cells grown in 3D are more physiologically relevant than cells cultured in 2D. To use 3D models in substance testing and regenerative medicine, reproducibility and standardization are important. Bioprinting offers not only automated standardizable processes but also the production of complex tissue-like structures in an additive manner. We developed an all-in-one  bioprinting  solution to produce soft tissue  models. The holistic approach included (1) a bioprinter in a sterile environment, (2) a light-induced bioink polymerization unit, (3) a user-friendly software, (4) the capability to print in standard labware for high-throughput screening, (5) cell-compatible inkjet-based printheads, (6) a cell-compatible ready-to-use BioInk, and (7) standard operating procedures. In a proof-of-concept study, skin as a reference  soft tissue model was printed. To produce dermal equivalents, primary  human dermal fibroblasts were printed in alternating layers with BioInk and cultured  for up to 7 weeks. During long-term cultures, the models were remodeled  and fully populated with viable and spreaded fibroblasts.  Primary  human dermal keratinocytes were seeded on top of dermal equivalents, and epidermis-like structures were formed as verified with hematoxylin and eosin staining and immunostaining. However, a fully stratified epidermis was not achieved. Nevertheless, this is one of the first reports of an integrative  bioprinting strategy for industrial routine application.

© 2015 Society for Laboratory Automation and Screening

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Figure Legend:

Bioprinting of a 3D skin model. A) Schematic representation of a printed dermal model. Alternating layers of BioInk™ (black spiral) and cell suspension (primary human dermal fibroblasts; orange spiral) were printed to produce a dermal equivalent. This structure was topped with circular BioInk™ layers serving as container for subsequent keratinocyte seeding. B) Top view of an entire printed dermal equivalent according to the scheme in A). The fibroblasts were cultured for 2 days and stained with MTT (viability staining; dark structures). The printing pattern was still visible.

Standardized 3D Bioprinting of Soft Tissue Models with Human Primary Cells. Global Medical Discovery

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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