RPTEC/TERT1

The RPTEC/TERT1 cells specifically respond to parathyroid hormone (PTH) but not arginine vasopressin (AVP), and react with enhanced ammonia genesis on lowering of the environmental pH.

The RPTEC/TERT1 cells exhibit sodium-dependent uptake of phosphate as well as intact functionality of the megalin/cubilin transport system.

RPTEC/TERT1 cells show the characteristic morphology and functional properties of normal proximal tubular epithelial cells.

At high cell densities, the RPTEC/TERT1 cells form characteristic "domes", maintain ultrastructural organization with tight junctions, densely packed microvilli and primary cilium, indicating functional cell polarization.

When cultured on the Corning™ Transwell™ Permeable membrane cell culture insert, the RPTEC/TERT1 cells at confluence form intact functional barrier as indicated by stabilized Trans-Epithelial Electrical Resistance (TEER) across the membrane.

1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

To make the complete growth medium, add hTERT RPTEC Growth Kit (ATCC^® ACS-4007™) to the base medium. The final concentration for each growth kit component in the complete hTERT immortalized RPTEC growth medium is as follows:  

• 5 pM triiodo-L-thyronine
• 10 ng/mL recombinant human EGF
• 3.5 µg/mL ascorbic acid
• 5.0 µg/mL human transferrin
• 5.0 µg/mL insulin
• 25 ng/mL prostaglandin E₁
• 25 ng/mL hydrocortisone
• 8.65 ng/mL sodium selenite
• 1.2 mg/mL sodium bicarbonate

Required but not supplied: G418 solution MUST be added to the above medium to a final concentration of 0.1 mg/mL G418 to maintain the selective pressure for immortalization

Note: Do not filter complete medium.

This medium is formulated for use with a 5% CO₂ in air atmosphere.

To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If storage of the frozen culture is necessary upon arrival, store the vial in liquid nitrogen vapor phase and NOT at -70°C. Storage at -70°C will result in loss of viability.

1. Prepare a 25 cm² or a 75 cm² culture flask containing the recommended complete culture medium. Prior to the addition of the vial contents, the vessel containing the growth medium should be placed in the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6) and to avoid excessive alkalinity of the medium during recovery of the cells.
2. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
3. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All operations from this point on should be carried out under strict aseptic conditions.
4. Transfer the vial contents to a centrifuge tube containing 9.0 mL of complete culture medium and centrifuge the cell suspension at approximately 250 x g  for 5 to 7 minutes. 
5. Discard the supernatant and resuspend the cells in fresh growth medium (see the batch-specific information for the recommended dilution ratio). Add this suspension to the prepared culture vessel.
6. Incubate the culture at 37°C in a suitable incubator. 
7. A 5% CO₂/95% air atmosphere is recommended if using the medium described on this product sheet.

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

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