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hTERT Immortalized RPTEC Growth Kit

ACS-4007

Growth kit to be added to DMEM:F12 (500 ml). The kit consists of individually packaged supplements; each supplement must be added in appropriate volumes as per Product Sheet directions to 500 ml basal medium, DMEM:F12.
Product category
Media
Comments
Each growth kit contains:
PCS-999-058 5 ml RPTEC Supplement A
PCS-999-059 10 ml RPTEC Supplement B

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 

The hTERT immortalized RPTEC Growth Kit contains components that when added to DMEM:F12 (ATCC® 30-2006™) create a complete cell culture environment for hTERT immortalized RPTEC cells (ATCC® CRL-4031™). The final concentration for each growth kit component in the complete hTERT immortalized RPTEC growth medium is as follows:
25 ng/mL Hydrocortisone
3.5 µg/mL Ascorbic Acid
8.65 ng/mL Sodium selenite
5.0 µg/mL Transferrin
5.0 µg/mL insulin
5 pM Triiodo-L-thyronine
25 ng/mL Prostaglandin E1
10 ng/mL rhEGF
1.2 mg/mL Sodium Bicarbonate 

Note: Fetal Bovine Serum is not required for proliferation but may be added if it is desired to transfect the cells (final FBS concentration in complete medium is 0.2%).

Product format
Frozen
Shipping information
Frozen supplement kit for 500 mL base medium
Storage conditions
-20°C or colder
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Documentation

ATCC determined that a biosafety level is not applicable to this material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to complete your own risk assessment and understand any potential hazards associated with the material per your organization’s policies and procedures and any other applicable regulations as enforced by your local or national agencies.

Detailed product information

General

Specific applications
Add to DMEM:F12 (ATCC® 30-2006™) to produce complete medium for hTERT Immortalized RPTEC cells (ATCC® CRL-4031™)

Handling information

Handling procedure

Unpacking and Storage Instructions

  1. Check all containers for leakage or breakage.
  2. Store the growth kit(s) at either -20°C in a freezer that is not self-defrosting or at -70°C for long term storage. If thawed upon arrival, the Growth Kit can be stored at 2°C to 8°C and added to the basal medium within 72 hours of receipt.

Preparation of Complete Growth Media

  1. Obtain one hTERT immortalized RPTEC Growth Kit from the freezer; make sure that the caps of all components are tight.
  2. Thaw the components of the growth kit and mix by gentle vortex or inversion just prior to adding them to the basal medium. Note: Only use 8.0 mL of Supplement B to obtain the optimal final concentration.
  3. Obtain one bottle of DMEM/F12 Medium (500 mL) from cold storage.
  4. Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying them with 70% ethanol.
  5. Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the indicated volume of each growth kit component, as indicated in Table 1, to the bottle of basal medium using a separate sterile pipette for each transfer.

Table 1. hTERT immortalized RPTEC Growth Kit

Component

Volume

Final Concentration

RPTEC Supplement A

5.0 ml

1% RPTEC Supplement A

RPTEC Supplement B

8.0 ml

1.6% RPTEC Supplement B

G418 (not supplied)

Varies based on source

0.1 mg/ml

 

The final concentration for each growth kit component in the complete hTERT immortalized RPTEC growth medium is as follows:

25 ng/mL Hydrocortisone

3.5 µg/mL Ascorbic Acid

8.65 ng/mL Sodium selenite

5.0 µg/mL Transferrin

5.0 µg/mL insulin

5 pM Triiodo-L-thyronine

25 ng/mL  Prostaglandin E1

10 ng/mL rhEGF

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.

Fetal Bovine Serum is not required for proliferation but may be added if it is desired to transfect the cells. The recommended volume of each optional component to be added to the complete growth media is summarized in Table 2.

Table 2. Addition of Serum  (Optional)

Component

Volume

Final Concentration

Fetal Bovine Serum,

1 ml

0.2% FBS

 

  1. Tightly cap the bottle of complete growth medium and swirl the contents gently to assure a homogeneous solution. Do not shake forcefully to avoid foaming. Label and date the bottle.
  2. Complete growth media should be stored in the dark at 2°C to 8°C (do not freeze). When stored under these conditions, complete growth media is stable for four weeks.

Quality control specifications

Bacterial and fungal testing
Not detected
Functional tests
Rate of proliferation and morphology comparable to control. Cells cultured in complete medium exhibit dome structures in 7-14 days

Legal disclaimers

Intended use
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.
Warranty

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.

Disclaimers

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.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Wieser M, et al. hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics. Am. J. Physiol. Renal Physiol. 295: 1365-1375, 2008. PubMed: 18715936

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