Water and electrolyte homeostasis is largely regulated by the human proximal tubule; however this complex system has been hard to study particularly in relation to specific genes and their influence on renal physiology and pathophysiology. Immortalized Human Proximal Tubule Cells (HPCT-03ts) is a potential model to acquire detailed physiological information about cell behaviour in response to hormones, growth factors, nutrients, and other external stimuli.
Immortalization Method: Immortalized with retrovirus containing temperature-sensitive SV40 large T antigen
BioSafety Level: II
Species: Human
Source Organ: Kidney
Growth Properties: Adherent
Morphology: Epithelial-cobblestone
Population Doubling: 24-48 hours
Seeding Density: 150,000 cells/cm²
Markers: NBCe1-A, NHE3, G418 resistance
Donor Age: 75 year old
Donor Gender: Female
Donor Ethnicity: N/A
Propagation: Cells are grown on Millicell Cell Culture Insert, 12 mm, hydrophilic PTFE, 0.4 µm (Millipore) that are lightly coated with a 20% dispersion of rat-tail collagen (in 60% ethanol) plus 1 µg/mL laminin.
The base medium for this cell line is Prigrow IV medium available at abm, Cat. No. TM004. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 5%, HEPES (Sigma) to a final concentration of 15 mM, sodium selenite (Sigma) to a final concentration of 20 nM, ascorbic acid-2-phosphate (Sigma) to a final concentration of 50 mM, transferrin (Sigma) to a final concentration of 5 µg/mL, Insulin Solution (TM053) to a final concentration of 5 µg/mL, mouse EGF (Sigma) to a final concentration of 10 ng/mL, dexamethasone to a final concentration of 4 µg/mL, T3 (Sigma) to a final concentration of 1 nM, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Change media every 2-3 days by first removing it from the bottom of the filter and then from the top. Add fresh medium first to the top and then to the bottom.
Carbon dioxide (CO2): 5%, Temperature: +33.0°C. To differentiate cells, increase temperature to +38.0°C.
* Do not use heat-inactivated FBS for cell culture unless specified otherwise.
Quality Control: 1) Western blot
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C