An in vitro model MPS model of human glomerulus for high throughput drug safety studies

抄録

The glomerulus is the structure in the kidney that filters waste products and small molecules from the blood into the urine, Essentially, the glomerulus is formed from an endothelial layer, a basement membrane and a size and charge selective sieve formed by podocytes. Damage to podocytes cells effectively destroys the size and charge selective filter, permitting the passage of larger and larger proteins from the blood into the urine, resulting in proteinuria. Currently there are no in vitro model of the glomerulus,

We have developed an in vitro 3D model of the glomerular filtration barrier (GFB), using human primary podocytes and endothelial cells derived from kidneys surplus to transplant requirements. We have developed a method for isolation of podocytes which yields a ~90% pure population. The podocytes express all known podocyte markers, including synaptopodin, WT1, nephrin, NEPH1, podocin, CD2AP, and podocalyxin. The podocytes also have central cilia, shown by pericentrin and acetylated tubulin staining. Cells are grown in cell culture inserts to generate an in vitro proteinuria assay. We have used this to demonstrate that our model retains the size selectivity of the in vivo GFB using a range of FITC-labelled dextran molecules of differing molecular weights and charges. We have shown that this size selectivity is disrupted using known podocyte toxins, adriamycin and puromycin. Our model is constructed in a 96-well plate format, making it amenable to high throughput analysis of GFB function and drug screening of compounds that affect GFB permeability and health.