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Globally, liver disease accounts for 2 million deaths per year of which cirrhosis, an advanced stage of fibrosis, causes 1.16 million deaths. Fibrosis is a response to chronic liver injury that can be initiated by several factors such as alcohol, non-alcohol fatty liver disease and viral hepatitis. LSECs play an important role in fibrosis and it is even suggested that LSECs could play a role in fibrosis initiation as the their specific phenotype changes (also known as capillarization) before fibrosis occurs in several liver diseases. LSECs line the sinusoidal wall and are characterized by the absence of a basal lamina and the  presence of open pores also known as fenestrae that act as a dynamic filter. During capillarization LSECs lose their fenestrations and obtain a basement membrane. Research aimed to elucidate mechanism involved in this capilarization of LSECs has focused on LSEC cultured in mono-layers whereby these cells capillarize due to the change to the artificial environment. Co-culture of LSECs with other liver cell types or conditioned medium of other liver cells can improve LSEC function in culture. However, to study LSECs in both healthy and diseased conditions, improvements need to be made for in vitro culture of LSECs. Therefore, we aim to develop a 3D spheroid culture with LSECs, hepatocytes and HSCs to unravel the morphology and diversity of LSECs in both healthy as diseased states. In particular, we are intrested to determine the health status of LSECs by the expression of functional markers and by conventional Electron Microscopy (EM) and Super Resolution Microscopy (SRM) that will be developed by the DeLIVER consortium.


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