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The E11 cell line is a highly specialized murine cell line developed for advanced studies in podocyte function and kidney disease mechanisms. Derived from the glomeruli of transgenic mice engineered to express a temperature-sensitive variant of the SV40 large T antigen, the E11 cells operate under the regulation of the IFN-g-inducible H-2kb promoter. This unique genetic framework facilitates the conditional proliferation of the cells, dependent on the environmental temperature, which aligns with the controlled expression of the T antigen.
One of the distinguishing features of the E11 cell line is its phenotypic stability across extensive passaging. Maintaining consistent expression and cellular characteristics through more than 40 passages, E11 cells have proven invaluable for long-term studies without the common issue of phenotypic drift seen in many cultured cell lines. This stability enhances their use in repetitive and extended biological experiments requiring consistent cell behavior.
In terms of protein expression, the E11 cell line exhibits a robust profile that is quintessential for podocyte-specific studies. The cells consistently express nephrin, an essential component of the slit diaphragm structure in podocytes, alongside a variety of other podocyte-specific proteins such as podocin, CD2AP, and synaptopodin. This comprehensive protein expression facilitates the study of podocyte biology in a controlled in vitro environment, closely simulating in vivo conditions. The ability of E11 cells to form extensive cell-cell contacts further underscores their suitability for modeling kidney filtration barrier functionalities.
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Protein expression: WT1, Lmx1b, nephrin, NEPHI, FAT, P-cadherin, CD2AP, ZO-I, podocalyxin, podoplanin, synpo, podocin, TRPC6 and GAPDH.
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: Inocculate T75 cell culture flasks with 1x 104 cells/cm2 for the proliferation process. Keep the cells at 33 degree Celsius / 5% CO2, until the flask is about 75% confluent.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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