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Time-dependent structural and functional characterization of subcutaneous human liver tissue.
Journal of Tissue Engineering and Regenerative Medicine 2018 October 24
Subcutaneous transplantation of engineered hepatocyte/fibroblast sheets (EHFSs) is a low invasive and safe approach to construct vascularized subcutaneous human liver tissue (VSLT). However, the liver-specific structures and functionalities in the development process of VSLTs in mice remain poorly understood. Here, we describe time-dependent characteristics of the formation of the vascular network, cell-cell adhesions, liver transporters, liver-specific protein synthesis, and metabolizing activities. The EHFSs formed multilayered thick tissues by rapid neovascularization, which allows overcoming extremely difficult problems, such as the lack of oxygen supply on the formation of three-dimensional primary hepatocyte tissue under the skin. The blood vessels consisted of mouse-origin endothelial cells (ECs) (mVEGFR2) from the subcutaneous space at 1-7 days, and the following formation of the vascular network was performed by human-origin ECs (hVEGFR2). Many varieties of liver-specific gene expressions increased with the construction of the VSLTs: cell-cell adhesion molecules (CDH1, CLDN3, and CX32), transporters at basal (OATP1A1, OCT1, and NTCP) and apical membranes (MRP2, MDR1, and BSEP), blood coagulation factors (F8 and F9), urea synthesis (CPS1, OTC, and ARG1), and metabolism enzymes (CYP7A1, CYP1A2, CYP2B6, CYP3A4, and UGT1A1). Subacute hepatic failure model mice with VSLT were alive at least 7 weeks after liver damage. Thus, the ectopic liver organ offers the potential for a low invasive and safe treatment for liver diseases.
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