S. Simonsson (Gothenburg, SE)
Institute of Biomedicine Department of Clinical Chemistry and Transfusion MedicinePresenter Of 1 Presentation
12.3.6 - 3D bioprinted meniscus tissue-like structure using gene edited iPS cell -line and chondrocytes induces the cartilage marker ACAN
Abstract
Purpose
Pathological changes of the meniscus are common in people with Osteoartrithis (OA) and damage to the meniscus often leads to secondary OA. Meniscus architecture is individual and a perfect match is suggested to be key for successful transplantation. For this reason and due to the limited availability of donor meniscus other strategies such as 3D bioprinting is sought for.
Methods and Materials
Herein we 3D bioprinted a meniscus prototype including human induced pluripotent stem cells (iPSCs) together with human chondrocytes, using nanocellulose/alginate containing bioink previously published. Chondrocytes were obtained from autologous chondrocyte implantation (ACI) surgery and the iPSC line was generated by reprogramming the chondrocytes. CRISPER-Cas9 gene editing of the chondrocyte derived iPSC line was used to insert the green fluorescence protein (GFP) under control of the aggrecan promoter (ACAN). After printing the meniscus prototypes were placed in chondrogenic-differentiation medium for 3 weeks. Sections of the 3D bioprinted meniscus were stained by Alcian Blue van Gieson stainining. The aggrecan expression (ACAN) analysis of the 3D bioprinted iPSC/Chondrocytes was performed after 3 weeks using fluorescence imaging microscope IN Cell Analyzer 6000 (IN Cell 6000, GE Healthcare, United Kingdom).
Results
Meniscus-like tissue structures were reproducible obtained and could be scaled down and printed in 96 well format (Figure). Proteoglycan staining supported differentiation along the fibrous-cartilage lineage similar to what is normally seen in meniscus native tissue. The cartilage marker ACAN was induced in the GFP positive cells so that they can be traced back to the gene edited iPSCs and darker areas were assumed to originate from the human chondrocytes. Vascular-like tubes could be seen in sections and the surrounding endothelial cells were developed from the iPSC population based on visualization of GFP expression.
Conclusion
In this study we were able to produce a meniscus structure by 3D bioprinting multiple meniscus prototypes resembling native meniscus tissue.