Immortalized Human Tonsillar Epithelial Cells (HTE)

Cat. No.
T0721
Unit
1x106 cells / 1.0 ml
Price
$710.00

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Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing institution.

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Cat. No. T0721
Name Immortalized Human Tonsillar Epithelial Cells (HTE)
Description

Immortalized Human Tonsillar Epithelial (HTE) cells are isolated from healthy adult patient biopsy and immortalized by being transfected with human papillomavirus type 16 (HPV16). HTE cells can be a useful cell line model as HPV-transfected human tonsillar epithelial cells for studying the HPV pathology and the role of HPV in the etiology of HPV related cancer. HTE cells can also be used as a cell model to study human tonsillar epithelial cell physiology and function.
Organism Human (H. sapiens)
Tissue Tonsil
Donor History Not disclosed
Growth Properties Adherent, polygonal
Unit 1x106 cells / 1.0 ml
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions Ship with dry ice.
Product Format Frozen
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.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions

For optimal cell culture, we recommend using PriCoat™ T25 Flasks (G299) or coating your preferred vessels with Applied Cell Extracellular Matrix (G422). PriGrow IX (TM019) + 5% FBS (Regular*) + 10 μM ROCK Inhibitor Y-27632 (TM131) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.

*Do not heat-inactivate

200 µg/ml Geneticin/G418 (G271) for selection.

Note: Selection drugs should be added to the culture medium after the first passage to ensure cells have recovered from freeze-thaw conditions.

Change media every 2-3 days once co-culture has been established.
Unpacking and Storage Instructions

1. Visually examine the packaging containers for signs of leakage or breakage.

2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.

To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.
Thawing Protocol

Feeder cells (NIH 3T3 J2 fibroblasts) must be prepared prior to thawing T0721. To prepare the feeder cell layer:

  1. Thaw NIH 3T3 J2 fibroblasts as per supplier instructions. Once fibroblasts reach 40-50% confluency, add Mitomycin C (15 μg/mL) and incubate at 37°C for three hours. 

  2. Remove mitomycin C containg growth media and wash cells for a total of 5X using PBS (1X) to remove any residual traces of mitomycin C. 

  3. Add fresh growth media to the culture vessel and incubate at 37°C (5% CO2) for 1-2 hours to allow cells to adapt. 

    Note: feeder cells may change morphology post-treatment with mitomycin C and some floating cells may be observed. Although cells may appear bigger in size and present multiple nuclei, confluency should not increase post-treatment. 

  4. The feeder lay is now ready for T0721.


Prepare T0721 for co-culture: 

  1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.

  2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.

  3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.

  4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media.


Co-culture T0721 and NIH 3T3 J2 fibroblasts feeder cells: 

  1. Aspirate feeder cell growth media from the culture vessel containing NIH 3T3 J2 fibroblasts feeder cells and replace with T0721 complete growth media.

  2. Add the re-suspended T0721 cells (from step 4 above) to the culture vessel and incubate cells according to the recommended culture conditions.  

  3. T0721 may take additional time to recover from post-thaw conditions, therefore, allow cells to attach over the next 24-48 hours before changing media. 

  4. A successful co-culture will present growing colonies of T0721 between the feeder cells. T0721 cells may grow on-top of one another. 
Subculture Protocol

Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.

  1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. Incubate at 37°C for one minute to remove feeder cell layer. 

  2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes) of only the feeder cells. Collect the suspended feeder cells and discard.

  3. Add 2-3ml of fresh, pre-warmed 0.25% Trypsin-EDTA to the culture vessel. Incubate at 37°C for 2-3 minutes. 

  4. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media to the culture vessel.

  5. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.

  6. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels that have a feeder cell layer ready. 

  7. Incubate the cells at the recommended conditions.
Cryopreservation We recommend using serum-free CryoGuard™  Freezing Media (TM078) or, if serum is preferred, Cryopreservation Medium (TM024).
Immortalization Method

Dual infection with E6 and E7 HPV16 viral genes.
Expression

E7 and E1^E4 expression; G418-resistant
Warranty abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period”.
Disclaimer

1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at licensing@abmgood.com.

2. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.

3. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).

4. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.

5. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.

6. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Depositor LSU Health Sciences Center - Shreveport
QC

1) Quantification of viral transcripts
Application Research Use Only.
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0721
Print/Download Datasheet

  • Spanos, W. C., Geiger, J., Anderson, M. E., Harris, G. F., Bossler, A. D., Smith, R. B., Klingelhutz, A. J., & Lee, J. H. (2008). Deletion of the PDZ motif of HPV16 E6 preventing immortalization and anchorage-independent growth in human tonsil epithelial cells. Head & neck, 30(2), 139–147. https://doi.org/10.1002/hed.20673


    Bienkowska-Haba, M., Luszczek, W., Myers, J. E., Keiffer, T. R., DiGiuseppe, S., Polk, P., Bodily, J. M., Scott, R. S., & Sapp, M. (2018). A new cell culture model to genetically dissect the complete human papillomavirus life cycle. PLoS pathogens, 14(3), e1006846. https://doi.org/10.1371/journal.ppat.1006846

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