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Development of renal podocytes cultured under medium perifusion
Background:
In the past, podocytes have been described as highly susceptible to dedifferentiation under cell culture. Whether this process resulted from insufficient culture conditions or whether it was a consequence of missing cellular interactions remained unclear. A further reason could be that podocytes within the maturing kidney are irreversibly growth-arrested at a very early point of development because proliferating cells have been detected at the S-shaped body stage but not at the capillary loop stage or in the maturing glomeruli. These were important reasons that hindered the establishment of podocyte cell culture systems.
Experimental Design:
The aim of our present study was to culture podocytes under the most organotypic conditions possible to maintain typical cellular characteristics. Cortex explants of neonatal rabbit kidneys consisting of nephrogenic tissue were used as a source for podocytes. No serum additives were given for the whole culture period of 13 days. An organ-specific environment was obtained by keeping the podocytes within the surrounding renal tissue and by ensuring a permanent exchange of medium.
Results:
mAb were used to characterize podocytes and the other glomerular cell types. Cultured podocytes and parietal cells of Bowman's capsule were identified by EnPo 1. Ks 19.2.105, a marker for cytokeratin 19, was used to discriminate among these epithelial cells because cytokeratin 19 is expressed by the parietal cells of Bowman's capsule but not by podocytes. The Ab EC1 specifically detected endothelial cells. Glomerular endothelium cultured under medium perifusion expressed these typical Ag and thus could be unequivocally discriminated. Furthermore, by means of the proliferation marker Ki-67, it could be demonstrated that glomerulus-like structures developed under culture by proliferation of visceral and parietal cells of Bowman's capsule.
Conclusions:
A culture model is presented that allows the maintenance of developing podocytes within the organ-specific tissue environment and under permanent medium perifusion.