Description: 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 antigenBioSafety Level: IISpecies: HumanSource Organ: KidneyGrowth Properties: AdherentMorphology: Epithelial-cobblestonePopulation Doubling: 24 - 48 hoursSeeding Density: 150,000 cells/cm10²Markers: NBCe1-A, NHE3, G418 resistanceDonor Age: 75 year oldDonor Gender: FemaleDonor Ethnicity: N/APropagation: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 blotShipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C