Generation of Patient-Derived Podocytes from Skin Biopsies

We describe a protocol for generating human podocytes by episomal reprogramming dermal fibroblasts into human-induced pluripotent stem cells. These cells resemble in vivo podocytes much better than immortalized podocytes in terms of podocyte food processes and expression of podocyte specific marker. Podocytes are terminally differentiated cells.

Therefore, these cells no longer proliferate and primary podocyte analysis in cell culture is limited. Even though the problem of limited cell number was overcome by using conditionally immortalized podocytes, these podocytes present a dedifferentiated phenotype and behavior, which questions their use in basic research. With the help of induced pluripotent stem cells, podocytes can be generated in vitro in an almost unlimited cell number with typical podocyte morphology including distinct food processes and expression of podocyte specific marker.

When using the patient-specific stem cells to differentiate into podocytes, it is possible to study the cell’s disease-specific alterations ex vivo. In case, the initial skin biopsy was taken from a patient with a genetic podocyte disease. Podocytes generated with our protocol are personalized diseased cells carrying the patient’s mutation.

Thus, these cells represent an improved ex vivo model to study podocyte diseases and potential therapeutic substances in an individualized approach.