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SVI
Tissue
Kidney

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SVI

Cell line · Product number 400495

The SVI cell line has been cloned from the outgrowth of glomeruli which were isolated from H-2kb-tsA58 transgenic mice. The mice carry a temperature-sensitiv...

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01General information

Description
The SVI cell line has been cloned from the outgrowth of glomeruli which were isolated from H-2kb-tsA58 transgenic mice. The mice carry a temperature-sensitive variant of the SV40 large T antigen under control of the IFN-g-inducible H-2kb promoter. Cells proliferate at 33 degree Celsius, and they differentiate at 37 degree Celsius. At present, the cells have been cultured successfully for more than 40 passages without noting phenotypic changes. SVI are very similar to E11 in terms of morphology and the expression of several markers. For example, podocin and WT1 are expressed to a lesser extent as compared to E11. Differentiation: Start the differentiation process by placing the non-confluent flask(s) into an incubator at 38 degree Celsius / 5% CO2 for a minimum of 14 days to complete the differentiation. Addition of interferon-gamma (INF-gamma) is not necessary.
Tissue
Kidney

02Classification

Organism
Mouse
Sex
Unspecified
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine podocyte cell line (SVI) contains a conditionally active SV40 Large T-Antigen transgene as part of the ImmortoMouse model, supporting temperature-sensitive immortalization. The construct is stably present in podocyte-derived cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Cell type
Podocyte

04Identifiers

Citation
SVI (Cytion catalog number 400495)
NCBI TaxID
10090
Cellosaurus
CVCL_5943

05Expression and susceptibility

Protein expression
WT1, Lmx1b, nephrin, NEPHI, FAT, P-cadherin, CD2AP, ZO-I, podocalyxin, podoplanin, synpo, podocin, TRPC6 and GAPDH.

06Handling and culture

Culture medium
RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
Incubation atmosphere

33°C, 5% CO2, humidified atmosphere.

Supplements
Supplement the medium with 10% FBS
Dissociation reagent
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.
Seeding density
Inocculate T75 cell culture flasks with 1x 104 cells/cm2 (about 60.000 cells/ml, 12ml medium in one T75) for the proliferation process. Keep the cells at 33 degree Celsius / 5% CO2, until the flask is about 75% confluent.
Fluid renewal
3 times per week
Thawing and culturing

  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

Freeze medium
As a cryopreservation medium, we 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.
Storage conditions

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Shipping conditions

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

08Donor

Breed / subspecies
(CBA/Ca x C57BL/10)Tg(H2KbtsA58) Immort
Donor age
Adult