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Primary Cervical Epithelial Cells

PCS-480-011

Product category
Human cells
Product type
Primary cell
Organism
Homo sapiens
Morphology
polygonal, cobblestone appearance
Tissue
Uterus; Cervix
Disease
Normal
Applications
3D cell culture
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

Biosafety Icon BSL 1

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.

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.

Cervical Epithelial Cell Basal Medium

PCS-480-032

Price: $66.00 ea

Cervical Epithelial Growth Kit

PCS-480-042

Price: $94.00 ea

Trypsin-EDTA for Primary Cells

PCS-999-003

Price: $28.00 ea

Trypsin Neutralizing Solution

PCS-999-004

Price: $28.00 ea

Dulbecco's Phosphate Buffered Saline (D-PBS), 1X

30-2200

Price: $16.00 ea

Detailed product information

General

Specific applications
Cervical epithelial cells provide an ideal in vitro model for studying the pathophysiology of cervical polyps, HPV, and cervical cancer.

Characteristics

Cells per vial
≥ 5.0 x 105
Volume
1.0 mL
Growth properties
Adherent
Age
lot-specific
Ethnicity
Lot-specific
Gender
Lot-specific
Comments
Characterization: Pan-Cytokeratin (+), TE-7 (-)

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

The basal medium for this primary cell is Cervical Epithelial Cell Basal Medium (ATCC PCS-480-032). To make the complete medium add the contents of Cervical Epithelial Cell Growth Kit (ATCC PCS-480-042) to the basal medium.

Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure
Refer to the batch specific information for the total number of viable cells recovered from this lot of ATCC® PCS-480-011™.
  1. Using the total number of viable cells, determine how much surface area can be inoculated to achieve an initial seeding density of 5000 cells per cm2.
  2. Prepare the desired combination of flasks. Add 5 mL of complete growth media per 25 cm2 of surface area. Place the flasks in a 37°C, 5% CO2, humidified incubator and allow the media to pre-equilibrate to temperature and pH for 30 minutes prior to adding cells.
  3. While the culture flasks equilibrate, remove one vial of PCS-480-011 from storage and thaw the cells 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 1 to 2 minutes).
  4. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by spraying with 70% ethanol. All operations from this point onward should be carried out under strict aseptic conditions.
  5. Add 5 mL of complete growth media – 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.
  6. Count the cells. Plate 5,000 cells per cm2 into each of the pre-equilibrated culture flasks prepared in steps 1 to 3 of Handling Procedure for Frozen Cells and Initiation of Culture. Gently rock the culture vessel from side to side and front to back to evenly distribute the cells within the vessel.
  7. Place the seeded culture flasks in the incubator at 37°C with a 5% CO2 atmosphere. Incubate for at least 48 hours before processing the cells further.
Subculturing procedure
  1. Passage normal cervical epithelial cells when culture has reached approximately 85 to 90% confluence, and are actively proliferating.
  2. Warm both the Trypsin-EDTA for Primary Cells (ATCC PCS-999-003) and the Trypsin Neutralizing Solution (ATCC PCS-999-004) to room temperature prior to dissociation. Warm complete growth medium to 37°C prior to use with the cells.
  3. For each flask, carefully aspirate the spent media without disturbing the monolayer.
  4. Briefly rinse the cell layer with 3 to 5 mL DPBS (ATCC 30-2200) to remove residual traces of serum and then aspirate and discard the DPBS.
  5. Add pre-warmed trypsin-EDTA solution (2 to 3 mL for every 25 cm2) to each flask.
  6. Gently rock each flask to ensure complete coverage of the trypsin-EDTA solution over the cells.
  7. Observe the cells under the microscope. When the cells pull away from each other and round up (typically within 3 to 5 minutes), remove the flask from the microscope and gently tap it from several sides to promote detachment of the cells from the flask surface.
  8. When the majority of cells are detached, quickly add an equal volume of Trypsin Neutralizing Solution (ATCC PCS-999-004) to each flask. Gently pipette or swirl the culture to ensure all of the trypsin-EDTA solution has been neutralized.
  9. Transfer the dissociated cells to a sterile centrifuge tube and set aside while processing any remaining cells in the flask.
  10. Add 3 to 5 mL Trypsin Neutralizing Solution to the flask to collect any remaining dissociated cells. Transfer remaining cells into the centrifuge tube.
  11. Repeat step 10 as needed until all cells have been collected from the flask.
  12. Centrifuge the cells at 270 x g for 3 to 5 minutes.
  13. Carefully aspirate the neutralized dissociation solution from the cell pellet and re-suspend the cells in 5 to 8 mL fresh, pre-warmed, complete growth medium.
  14. Count the cells and seed new flasks at a density of 5,000 cells per cm2.
  15. Place freshly seeded flasks in a 37°C, 5% CO2 incubator for at least 24 to 48 hours before processing the cells further.

Every alternate day, remove medium and feed 5 mL of supplemented medium. However, when cultures reach 50% (or greater) confluence, remove medium and feed with 5 to 8 mL of supplemented medium daily.

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Virus testing
Hepatitis B virus (HBV): Not detected
Hepatitis C virus (HCV): Not detected
Human immunodeficiency virus 1 (HIV-1): Not detected
Human immunodeficiency virus 2 (HIV-2): Not detected
Functional tests
Pan-Cytokeratin (+), TE-7 (-)
Population doubling capacity
When seeded at 5,000 cells per cm2, cells reach 95% confluence in 4 to 6 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

Woodworth CD, et al. Interleukin 1 alpha and tumor necrosis factor alpha stimulate autocrine amphiregulin expression and proliferation of human papillomavirus-immortalized and carcinoma-derived cervical epithelial cells. PNAS 92(7): 2840-844, 1995. PubMed: 7708734

Pecoraro G, et al. Differential effects of human papillomavirus type 6, 16, and 18 DNAs on immortalization and transformation of human cervical epithelial cells PNAS 86(2): 563-567, 1989. PubMed: 2463631

Ye F, et al. Stem-cell-abundant proteins Nanog, Nucleostemin and Musashi1 are highly expressed in malignant cervical epithelial cells. BMC Cancer 8: 108, 2008. PubMed: 18419830

Frequently Asked Questions

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