Immortalized Human Glomerular Podocytes-RFP-SV40 (Tet-on)

Immortalized human glomerular podocytes-RFP-SV40 (Tet-on) are conditionally immortalized human podocytes engineered with a doxycycline-inducible (Tet-on) SV40 large T antigen and stable red fluorescent protein (RFP) expression. In the presence of doxycycline, T antigen drives proliferation, enabling scalable expansion and reliable cryopreservation. Upon doxycycline withdrawal, proliferation ceases and cells differentiate into a mature podocyte-like phenotype, expressing nephrin, podocin, CD2AP, WT1, synaptopodin, podocalyxin, α-actinin-4, and ZO-1, with an arborized actin cytoskeleton and slit-diaphragm-associated structures.

The key advantage is dual control and traceability. Tet-on regulation allows temporal control of immortalization: expansion when needed, then differentiation with reduced T-antigen interference. Constitutive RFP enables non-invasive live-cell imaging, fluorescence-activated cell sorting, accurate quantification, and tracking in co-culture or 3D systems, including glomerular endothelial cell and mesangial cell models.

Compared with constitutive SV40 lines, these cells better support physiological studies while retaining human origin, batch consistency, and reduced donor variability. They are widely used to model podocyte injury, proteinuria, nephrotic syndrome, diabetic nephropathy, focal segmental glomerulosclerosis, and drug/toxin nephrotoxicity. They support mechanistic studies of insulin signaling, autophagy, apoptosis, cytoskeletal remodeling, and glomerular crosstalk, and are compatible with siRNA/CRISPR, immunostaining, qPCR, Western blot, and permeability assays.