PODO/TERT2562021-08-02T06:59:08+00:00

Kidney

PODO/TERT256

Evercyte ́s human kidney-derived podocytes cell line PODO/TERT256 can be grown without limitations while maintaining expression of cell type specific markers and function. Therefore, these cells are valuable as standardized in vitro model to study kidney diseases such as glomerulopathies. Moreover, the cells are useful for toxicity studies in pre-clinical drug development as well as for the development of glomerulus-protective therapeutics.

General information

Cat#: CHT-033-0256

Organism: homo sapiens
Tissue, cell type: kidney tissue (female), podocytes, visceral epithelial cells
Morphology:
typical podocyte morphology, cytoplasmic extensions partially with arborized appearance
Life span extension:
ectopic expression of hTERT
Quality:
free from contaminations (bacteria incl. mycoplasma, fungi, HIV, HAV, HBV, HCV, Parvo-B19) and cross-contaminations

Morphology and growth

PODO/TERT256 cells are characterized by the typical, podocyte morphology.The cells can be grown continuously for at least 80 PDs without showing signs of growth retardation.

Expression of marker proteins

PODO/TERT256 cells express typical podocyte marker proteins such as Nephrin, WT-1 and Synaptopodin. Staining of actin fibers further underlines the typical podocyte morphology. Cell nuclei are counterstained with DAPI.

PAN induced toxicity – 2D

Undifferentiated and differentiated PODO/TERT256 cells grown in multi-well plates were treated with puromycin aminonucleoside. Using viability assay we could demonstrate that the cells loose viability and that this PAN induced toxicity could in part be rescued by addition of Mizoribin (MRZ).

PAN induced toxicity / 3D co-culture

PODO/TERT256 cells were co-cultured with telomerised microvascular endothelial cells HDMVEC/TERT164 in transwells. The thereby formed barrier retains bovine serum albumin (BSA), but can be disrupted by PAN. Mizoribin (MRZ) can rescue PAN induced injury.

FAQs

What is the difference between PODO/TERT256 and PODO/SVTERT152 cells?2021-06-21T09:29:19+00:00

PODO/TERT256 cells, isolated from kidney tissue biopsies have been immortalized by reactivation of telomerase alone, whereas urine-derived PODO/SVTERT152 cells overexpress the catalytic subunit of human telomerase together with Simian Virus 40 largeT antigen. Both cell lines show typical markers of podocytes.

Is it necessary to cultivate the cells in the presence of G418?2021-06-21T09:28:28+00:00

G418 was used to select for stable positively transfected cells. Even if we have not systematically tested the stability of PODO/TERT256 cells when cultivated without G418, we assume that the addition of G418 is not necessary. However, after thawing the original ampoule, we recommend setting up a cell bank before testing the stability of the cells in medium without G418.

In vitro propagation

PodoUp3 ready-to use medium (Evercyte, Cat# MHT-033-3)

MCDB131 (Pan Biotech, Cat# P04-80057)

1,6 mM GlutaMAX-I (Gibco, Cat# 35050-038)

9,6 µg/mL bovine brain extract (Lonza, Cat# CC-4098)

8 ng/ml hEGF (Sigma Aldrich, Cat# E9644)

20 ng/ml Hydrocortisone (Sigma Aldrich, Cat# H0396)

20% fetal bovine serum (Sigma Aldrich, Cat# F7524)

100 µg/ml G418 (InvivoGen, Cat# ant-gn-5)

Additional material & reagents

Phosphate buffered saline (PBS) (Sigma, Cat# D8537)

50 µg/ml collagen I (Sigma Aldrich, Cat# C2249), diluted in PBS

0,05 % Trypsin-EDTA (Gibco, Cat#25300-054)

Protocol passaging of PODO/TERT256
Passaging of cells

The new culture flasks have to be pre-coated with human collagen I. Therefore, the culture flasks are pre-treated with collagen I solution (80 µl/cm²) at 37°C for at least 30 min.

Before introducing cells, remove excess of collagen I solution and rinse flask once with PBS (160 µl/cm²). For detachment of the cells remove and discard the culture medium and wash the cells twice with PBS. Remove PBS completely.
Then, add 0.05% Trypsin-EDTA (1x) solution (20 µl/cm²), make sure that all cells have been in contact with this solution and incubate the culture flask at 37°C for approximately 3 – 4 min. Observe cell detachment under an inverted microscope.
As soon as all cells are detached (if necessary, agitate the cells by gently hitting the flask), add growth medium (about 160 µl/cm²) and aspirate cells by pipetting.
Add appropriate aliquots of the cell suspension to collagen I pre-coated culture vessels supplemented with growth medium (final volume of 240 µl/cm²).
Cells should be split every 3-4 days (after having reached not more that 80 % confluence) with a split ratio of 1:4 to 1:6. Never allow the culture to become confluent!

Cryopreservation

Freezing medium

PodoUp3 medium (Evercyte, Cat# MHT-033-3)

10 % DMSO (Sigma Aldrich, Cat# D2650)

Additional material & reagents

Phosphate buffered saline (PBS) (Sigma, Cat# D8537)

50 µg/ml collagen I (Sigma Aldrich, Cat# C2249), diluted in PBS

0,05 % Trypsin-EDTA (Gibco, Cat#25300-054)

Protocol cryopreservation of PODO/TERT256
Freezing of cells

Detach the cells from the culture vessel by using Trypsin-EDTA solution (Protocol passaging of PODO/TERT256).

Resuspend the detached cells in growth medium and centrifuge at 170 g for 5 min.
Discard the supernatant, resuspend in the remaining droplet and add freezing medium (tempered to 4°C) to reach a cell density of about 7 x 10e5 cells /ml (for thawing in a 25 cm² culture flask).
Add 1 ml of this cell suspension to each pre-cooled cryovial and immediately transfer the cells to -80°C.
After 24 hours transfer the vials to the liquid nitrogen tank.
Thawing of cells

Original Evercyte cells are to be thawed in a T25 rouxflask

Pre-coat a 25 cm² culture flask with collagen I (Protocol passaging of PODO/TERT256).
Add 6 ml of growth medium to the pre-coated 25 cm² culture flask and place the culture flask in the incubator for at least 20 min to allow the medium to reach its normal pH.
Take a vial of frozen cells, rinse it outside with Ethanol and pre-warm in hand until one last piece of frozen cells is seen.
Then, immediately transfer the content of the vial to a 15 ml centrifugation tube pre-filled with 9 ml of medium pre-cooled to 4°C and centrifuge for 5 min at 170 g.
Discard the supernatant and resuspend the cell pellet in the remaining droplet.
Add 1 ml of pre-warmed medium to the cell suspension, transfer the cells to the prepared culture flask and incubate at 37°C in a suitable incubator.
Perform a medium change 24 hours after thawing. If the cells are already 70-80 % confluent at this point, they have to be passaged.
Protocol passaging of PODO/TERT256

Product data sheet – certificate of analysis

Product data sheet (PDS) download
Certificate of analysis

is available upon request | Please contact us indicating the respective LOT numbers

 

Protocols

Preparation of cell culture medium PodoUp3

Protocol passaging of PODO/TERT256
Protocol cryopreservation of PODO/TERT256
Protocol for 2D differentiation of PODO/TERT256

Data on Markers and Functions

PODO/TERT256 – morphology and growth characteristics
PODO/TERT256 –expression of nephrin, synaptopodin, WT-1 and F-actin
PODO/TERT256 – puromycin aminonucleoside (PAN) induced toxicity / 2D
PODO/TERT256 – puromycin aminonucleoside (PAN) induced toxicity / 3D

Safety documents (coming soon)

Telomerized human cells – material safety data sheet
PODO/TERT256 – cell line establishment, vector maps

 

Selected publications

Stevens M, et al. (2019) The natural drug DIAVIT is protective in a type II mouse model of diabetic nephropathy. PLoS One. 2019; 14(3): e0212910. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6415805/

Licence Conditions

The business concept of Evercyte is to out-license telomerized cells to our customers. The license conditions depend on whether the contract partner is a for profit or a nonprofit organization and the intended use of the cells.

Nonprofit organizations

Evercyte grants licenses for an unlimited period to academic or nonprofit-organizations, whereby the use of Evercyte cell lines is restricted to research & development purposes and non-commercial use. The cells are not intended for human use.
The customers have to agree to the conditions described in our material transfer agreement as well as accept our general terms and conditions.
On time payment for unlimited use: EUR 1300

Profit organizations

Pharmaceutical – chemical- cosmetic industries
Evercyte grants licenses for commercial organizations, whereby we offer an initial testing phase for a flat fee that allows our customers to test our cells in their laboratories for a period of 6 months.
Thereafter, annual license fees fall due, depending on the cell line of interest. Besides offering cell lines for research & development purposes, we also have established cell factories that qualify for production of clinical grade extracellular vesicles for human application.
The customer has to agree to the conditions described in our license agreements.
Contract research organizations (CRO)
Evercyte grants licenses for contract research organizations, whereby we offer an initial testing phase for a flat fee that allows our customers to test our cells in their laboratories. Thereafter, we would negotiate a royalty based long-term license agreement individually.
The use of the cells during these phases is restricted to research & development purposes. The cells are not intended for human use. The customers have to agree to the conditions described in our material transfer agreement and accept our general terms and conditions.
Initial license fee for 6 months: EUR 2000
Annual license fee R&D: royalty based
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Cat#: CHT-033-0256

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Customer Reviews

“I have had the pleasure of working with Evercyte for the last few years. We continually rely on Evercyte because of the high-quality data that they produce, their diligent responsiveness, and their excellent customer service.”

 

Josh Garlich, Senior Research Scientist, Apellis Pharmaceuticals, Inc.

“Cytonus has been working with Evercyte from many years as they are a trusted partner and have always delivered the highest quality cell lines to advance our platform.  We routinely draw on their expertise to meet cellular engineering challenges and they have not disappointed.”

Remo Moomiaie-Qajar, Cytonus Therapeutics, Inc.

Good cell culture practice / GCCP

get ready to standardize your cell culture work

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urine-derived podocytes

PODO/SVTERT152 cells from healthy donor

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PodpUp-3

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specific request

Customer Reviews

“I have had the pleasure of working with Evercyte for the last few years. We continually rely on Evercyte because of the high-quality data that they produce, their diligent responsiveness, and their excellent customer service.”

 

Josh Garlich, Senior Research Scientist, Apellis Pharmaceuticals, Inc.

“Cytonus has been working with Evercyte from many years as they are a trusted partner and have always delivered the highest quality cell lines to advance our platform.  We routinely draw on their expertise to meet cellular engineering challenges and they have not disappointed.”

Remo Moomiaie-Qajar, Cytonus Therapeutics, Inc.

Print Friendly, PDF & Email

from

€ 1300,–

PODO/TERT256
Cat#: CHT-033-0256

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO

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