Primary Cervical Epithelial Cells (ATCC® PCS-480-011)

Organism: Homo sapiens, human  /  Tissue: cervix  /  Disease: normal

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Organism Homo sapiens, human
Tissue cervix
Product Format frozen 1.0 mL
Morphology polygonal, cobblestone appearance
Culture Properties adherent
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease normal
Age lot-specific
Gender lot-specific
Ethnicity lot-specific
Applications Cervical epithelial cells provide an ideal in vitro model for studying the pathophysiology of cervical polyps, HPV, and cervical cancer.
Storage Conditions liquid nitrogen vapor phase
Images
Comments Characterization: Pan-Cytokeratin (+), TE-7 (-)
Complete Growth 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.

Subculturing
  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.

Culture Conditions
Atmosphere: air, 5%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial ≥ 5.0 x 105 viable cells
Volume 1.0 mL
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viral Testing Hepatitis B: None detected
Hepatitis C: None detected
Human immunodeficiency virus 1: None detected
Human immunodeficiency virus 2: None detected
Population Doubling Time When seeded at 5,000 cells per cm2, cells reach 95% confluence in 4 to 6 days.
Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
FAQ's
  1. Primary Cervical Epithelial Cells - PCS-480-011 passages

    HCxECs can be passaged over 4 passages (>10 PDLs).


    Date Updated: 7/22/2019
  2. Primary Cervical Epithelial Cells - PCS-480-011 subculturing

    It is recommended to passage HCxECs whey they reach ~85%  confluency.


    Date Updated: 7/22/2019
  3. Primary Cervical Epithelial Cells (HCxECs) (ATCC® PCS-480-011™) markers

    ATCC has tested HCxECs for positive expression of CK8, and CK18 markers and negative expression of TE-7 and vWF via ICC staining.


    Date Updated: 7/22/2019
  4. Primary Cervical Epithelial Cells - PCS-480-011 seeding density


    Date Updated: 7/22/2019

References

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