Immunohistochemistry confirmed retention of proteins in the scaffolds. The resulting scaffolds were validated using THLE-3 hepatocytes. We combined decellularized human liver tissue with electrospun polymers to produce a niche for hepatocytes and compared the human liver ECM to its individual components Collagen I, Laminin-521 and Fibronectin. Enhancing microenvironments using bioactive molecules allows researchers to create more appropriate niches for hepatocytes. Polymers and decellularized tissue scaffolds each provide some of the necessary biological cues for hepatocytes, however, neither alone has proved sufficient. One of the challenges for tissue engineers is the extracellular matrix (ECM) a finely controlled in vivo niche which supports hepatocytes. The results combined of this project are promising for future research, especially regarding the use of liver-derived organoids for recellularization.Tissue engineering of a transplantable liver could provide an alternative to donor livers for transplant, solving the problem of escalating donor shortages. The qPCR data showed variability between the different organoid lines and between the different phases of the organoid culture. The liver-derived organoids were successfully cultured and expanded in spinnerflasks, and were found engrafted and alive after 10 days in the scaffold. The HepG2 cell line validated the set up and recellularization with the HA, the cells engrafted throughout the scaffold and showed viability and signs of proliferation. The experiments were analyzed by histological and immunochemical staining and by qPCR. A setback in the project was the proneness to infections in the Harvard Apparatus (HA), which shortened the duration of experiments and influenced the results. In the bioreactor, the Harvard Apparatus (Hugo Sachs Elektronik), decellularized porcine liver segments were infused with a HepG2 cell line and liver-derived organoids in seperate experiments. This project shows that it is possible to use a perfusion-based bioreactor for repopulating a porcine liver scaffold with liver-derived organoids. However, there is no optimal method yet to recellularize such a decellularized liver scaffold. This ECM provides the biophysical and biochemical cues needed for cells to adhere, proliferate and differentiate. Luckily, the decellularization of an organ creates a scaffold that consists of the extracellular matrix (ECM) with important growth factors, bifunctional molecules such a fibronectin and multiple collagen types. Engineering organs with a complex structure and large vascular network, such as the liver, remain a challenge. Over the last decade new methods are explored in the field of tissue engineering to minimize the donor organ shortage. Vermeulen, Annewiet (TU Delft Mechanical, Maritime and Materials Engineering)īiomedical Engineering | Biomaterials and Tissue Biomechanics Repopulating a decellularized liver scaffold with liver-derived organoids in a perfusion-based bioreactor
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