Biochemistry, Department of

 

First Advisor

Edward N. Harris

Date of this Version

12-2020

Citation

A dissertation presented to the faculty of the Graduate College at the University of Nebraska in partial fulfillment of requirements for the degree of the Doctor of Philosophy

Major: Biochemistry

Under the supervision of Professor Edward N. Harris

Lincoln, Nebraska, December 2020

Comments

Copyright 2020, Fatima Cabral

Abstract

Current literature described techniques for the purification of liver cell types through text alone. The techniques described for the isolation of liver sinusoidal endothelial cells as well as hepatocytes described here are modified from a published article in the Journal of Visualized experiments. The video protocol allows for the user to successfully isolate cells as the most difficult parts of the procedure are demonstrated visually. The detection of liver maladies such as the non-alcoholic fatty liver disease, the stage if this disease and differentiation between non-alcoholic and alcoholic fatty liver disease is demonstrated in the development of a unique panel of protein kinase chemosensors. These have been previously developed to fingerprint mitogen activated protein kinase profiles. The liver has a unique subset of endothelial cells, as they are the only endothelial cell subtype that does not contain a basement membrane. This allows for ease of passage of blood-borne lipids and proteins into the parenchymal cells of the liver for metabolism and secretion. The liver sinusoidal cells also contain a host of receptor proteins including Stabilin-1 and Stabilin-2. Here we provide evidence that the Stabilin protein receptors provide a significant role in the rapid clearance of lipopolysaccharides from the blood. This is provided in blood clearance assays, as well as endocytosis assays in isolated liver sinusoidal endothelial cells derived from Stabilin knock out mice.

Advisor: Edward N. Harris

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