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Primary Gingival Keratinocytes


Primary gingival keratinocytes were isolated from the jaw and have important applications in antibiotic treatment, dental implants, and many other applications for oral biology research.
Product category
Human cells
Product type
Primary cell
Homo sapiens, human
Cell type
epithelial-like; cobblestone appearance; cells are rounded, not flat; cells display a high mitotic index; at near 80% confluence, the cells will be associated with each other in colonies.
3D cell culture
Product format
Storage conditions
Vapor phase of liquid nitrogen
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ATCC determines the biosafety level of a 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 understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

All tissues used for isolation are obtained under informed consent and conform to HIPAA regulations to protect the privacy of the donor’s Personally Identifiable Information. It is best to use caution when handling any human cells. We recommend that all human cells be accorded the same level of biosafety consideration as cells known to carry Human immunodeficiency virus (HIV) and other bloodborne pathogens. With infectious virus assays or viral antigen assays, even a negative test result may not exclude the possibility of the existence of a latent viral genome or infectious viral particles below the lower limit of detection of that assay.

ATCC recommends that appropriate safety procedures be used when handling all primary cells and cell lines, especially those derived from human or other primate material. Handle as a potentially biohazardous material using universal precautions. Cells derived from primate lymphoid tissue may fall under the regulations of 29 CFR 1910.1030 Bloodborne Pathogens.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information


Specific applications
The normal gingival epithelial cells, like the dermal epithelial cells express different types of anti-microbial peptides. Epithelial antimicrobial peptides play an important role in determining the outcome of the host-pathogen interaction at the oral mucosal barrier. Gingival keratinocytes may have important applications in antibiotic treatment, dental implants, and many other applications for oral biology research.


Cells per vial
Approximately 5.0 x 105
1.0 mL
Growth properties
Cryopreserved human oral keratinocytes derived from adult gingival tissue. Oral keratinocytes are cryopreserved in P2: After isolation from tissue the cells are cultured for two passages and then cryopreserved. Vials contain at least 500,000 cells in one mL cryopreservation solution.

Handling information

Unpacking and storage instructions
  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.
Complete medium
One bottle of Dermal Cell Basal Medium (ATCC® PCS-200-030™) plus one Keratinocyte Growth Kit (ATCC® PCS-200-040™).
95% Air, 5% CO2
Handling procedure

1. Refer to the batch specific information for the total number of viable cells recovered from this lot of ATCC PCS­-200­-014.

2. Using the total number of viable cells, determine how much surface area can be inoculated to achieve an initial seeding density of 5,000 cells per cm2.

3. Prepare the desired combination of flasks. Add 5 mL of complete growth medium per 25 cm2 of surface area. Place the flasks in a 37°C, 5% CO2 humidified incubator and allow the media to preequilibrate to temperature and pH for 30 minutes prior to adding cells.

4. While the culture flasks equilibrate, remove one vial of ATCC PCS-­200-­014 from storage and thaw the cells by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 1 to 2 minutes).

5. 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 onward should be carried out under strict aseptic conditions.

6. Add the appropriate volume of complete growth medium [volume = (1 mL x number of flasks to be seeded) ­ 1 mL] into a sterile conical tube. Using a sterile pipette, transfer the cells from the cryovial to the conical tube. Gently pipette the cells to homogenize the suspension. Do not centrifuge.

7. Transfer 1.0 mL of the cell suspension to each of the pre­equilibrated culture flasks prepared in steps 1 to 3 of Handling Procedure for Frozen Cells and Initiation of Culture. Pipette several times, then cap and gently rock each flask to evenly distribute the cells.

8. Place the seeded culture flasks in the incubator at 37°C, 5% CO2 atmosphere. Incubate for at least 24 hours before processing the cells further.

Subculturing procedure
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Remove and discard culture medium.
  2. Rinse the cell layer with DPBS solution for 2 minutes to remove all traces of serum that contains trypsin inhibitor.
  3. Add 5.0 to 7.0 mL of Trypsin-EDTA solution to the flask and incubate at 37°C .Observe cells under an inverted microscope until cell layer is dispersed (usually within 4 to 6 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Add 5.0 to 7.0 mL of Trypsin Neutralizing Solution (ATCC® PCS-999-004™) Centrifuge at 125 x g; 10 ± 2 minutes. Discard supernatant and resuspend the cell pellet with 8 mL of complete growth media. Gently break cell pellet by pipetting repeatedly.
  4. Count cells. Seed 2,500 to 5,000 viable cells per cm2. Add appropriate volume of the cell suspension to new culture vessels.
  5. Incubate cultures at 37°C.
Change media every 2-3 days
Subculture when cells reach 75-80% confluence. Seeding density should be 2,500 to 5,000 viable cells per cm2
Culture maintenance

The flask was seeded with cells (see specific batch information) grown and completely filled with medium at ATCC to prevent loss of cells during shipping.

  1. Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination. Using an inverted microscope (preferably equipped with phase-contrast optics), carefully check for any evidence of microbial contamination.  Also check to determine if the majority of cells are still attached to the bottom of the flask; during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable).
  2. If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium.  The shipping medium can be saved for reuse.  Incubate the cells at 37°C in a 5% CO in air atmosphere until they are ready to be subcultured.
  3. If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 125 x g for 5 to 10 minutes.  Remove shipping medium and save.  Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm2 flask.  Incubate at 37°C in a 5% CO2  in air atmosphere until cells are ready to be subcultured.

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Virus testing
Human immunodeficiency virus 1 (HIV-1): Not detected
Hepatitis C virus (HCV): Not detected
Hepatitis B virus (HBV): Not detected
Human immunodeficiency virus 2 (HIV-2): Not detected
Population doubling capacity
15 PDL
≥ 70% when thawed from cryopreservation


Year of origin

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.

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.

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

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.


Frequently Asked Questions


Curated Citations

Hou C, et al. MircoRNA-509 acts as a tumor suppressor in tongue squamous cell carcinoma by targeting epidermal growth factor receptor. Mol Med Rep 16(5):7245-7252, 2017. PubMed: 28944863

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