Aim 1:
To expand and unify the knowledge of 3D in vivo architecture of the developing pancreas
We will generate systematic information documenting the 3D pancreatic tissue organization and topological relationships of the epithelium with the surrounding mesenchyme and vasculature.
Gene expression information (bulk and single-cell RNASeq datasets) of the three cell populations will be integrated with detailed image reconstruction (light-sheet microscopy datasets) of the 3D organization of the epithelium, mesenchyme and vascular network and their topological relationships in the developing pancreas. Moreover, quantitative data on epithelial, mesenchymal and endothelial cell abundance will be collected. Such a deep understanding of the multi-cellular and multi-scale organization of the developing pancreas, including cell-cell interactions and gene expression profiling, will serve to generate computer-based 3D models for bioprinting applications.
Schematic representation of pancreas tissue development (top) and bioprinting approach (bottom). (a)Light-sheet fluorescence microscopy (LSFM) of E18.5 mouse pancreas. (b)(c) Immunofluorescent staining showing the multicellular composition of embryonic pancreas. mes, mesenchyme; EC, endothelial cells.
Aim 2:
To develop bioprinting solutions for engineering pancreatic tissue units
We have built an interdisciplinary team that will work together to develop a unique approach for the biofabrication of pancreatic modules that recapitulate native pancreatic tissue-architecture.
Specifically, the 3D bioprinting company Poietis together with the academic groups will 1) define appropriate bioprinting conditions for each cell type; 2) define appropriate biomaterials, which are compatible with living cells and printing procedures, and provide mechanical as well as functional properties (e.g. ECM and bioactives); 3) provide well-characterized cellular sources, including pancreatic progenitors and matching mesenchymal populations. Moreover, efforts will be targeted to build cellular constructs that are fully vascularized, which is a current roadblock in the field.
Aim 3:
To establish conditions for in vitro differentiation and maturation of the bioprinted pancreatic tissue
Our aim here is to establish conditions for in vitro differentiation and maturation of the bioprinted pancreatic tissue and to extensively study the tissue after the organ-printing process.
Engineered tissues will provide ex vivo models for studying stem cell and developmental biology, physiology, diseases of the pancreas, and drug toxicity. Over the long term, they will serve as guidelines to engineer pancreas for transplantation and regenerative medicine. A prerequisite to attain these final objectives is that the bioprinted tissues exhibit mature phenotypes and functions. We will systematically define critical factors and culture conditions capable of supporting either pancreatic progenitor maintenance and expansion or maturation into endocrine or exocrine tissue units.