Transplantation of patient-specific bile duct bioengineered with chemically reprogrammed and microtopographically differentiated cells
Cholangiopathy is a diverse spectrum of chronic progressive bile duct disorders with limited treatment options and dismal outcome. Scaffold- and stem cell-based tissue engineering technologies hold great promise for the reconstructive surgery and tissue repair. Here, we report a combined application of 3D scaffold fabrication and direct reprogramming of the patient-specific human hepatocytes to a population of pluripotent stem cells to fabricate implantable artificial tissues that imitate mechanical and biological properties of the native bile ducts. The chemically derived hepatic progenitor cells (hCdHs) were generated using two small molecules A8301 and CHIR99021 and seeded inside the tubular scaffold engineered as synergistic combination of two layers. The inner electrospun fibrous layer was made of nanoscale-macroscale polycaprolactone fibers acting to promote the hCdHs attachment, alignment, and differentiation, while the outer microporous acellular foam layer served to increase mechanical stability. The two layers of fiber and foam were fused robustly together thus creating a coordinated mechanical flexibility to exclude any possible breaking or tear during surgery. The gene expression profiling and histochemical assessment confirmed that hCdHs acquired the biliary epithelial phenotype and filled the entire volume surface of fibrous matrix after two weeks of growth in cholangiocyte differentiation medium in vitro. The fabricated construct replaced the macroscopic part of the common bile duct (CBD) and re-stored the bile flow in a rabbit model of acute CBD injury. Animals which received the acellular scaffolds did not survive the replacement surgery. Thus, the artificial bile duct constructs populated with the patient-specific hepatic progenitor cells could provide a scalable and compatible platform for treating bile duct diseases. Overall design: Transcriptome analysis for chloangiocytes differentiation of human chemically induced hepatic stem cells (hCdHs)