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hTERT-HPNE E6/E7; Pancreatic Duct; Human (Homo sapiens) Item ID: ATCCRL4036

Permits and Restrictions

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Organism Homo sapiens, human
Tissue pancreas, duct
Product Format frozen
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 2  [Cells contain Human Papilloma (HPV16) viral DNA sequences]

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.
Age 52 years
Gender male
Storage Conditions liquid nitrogen vapor phase
Karyotype This is a cell line of male origin that contains two major clonal cell populations: 45~47, XY,der(21)t(17;21)(q21.3;p13) and 46,XY, t(3;18) (p21.1;q11.2),der(21)t(17;21)(q21.3;p13). Other chromosomal aberrations were observed in the examined cells of both clones, but none were of a consistent nature.
Images
Clinical Data male
Antigen Expression positive for nestin (flow cytometry) (verified at ATCC)
Genes Expressed positive for nestin (flow cytometry) (verified at ATCC)
Tumorigenic NO
Complete Growth Medium The base medium for this cell line is:
  • 75% DMEM without glucose (Sigma Cat#. D-5030 with additional 2 mM L-glutamine and 1.5 g/L sodium bicarbonate)
  • 25% Medium M3 Base (Incell Corp. Cat# M300F- 500)
To make the complete growth medium, add the following components to the base medium:
  • fetal bovine serum 5% (final conc.)
  • 10 ng/ml human recombinant EGF
  • 5.5 mM D-glucose (1g/L)
  • 750 ng/ml puromycin
Subculturing Protocol: Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37.0°C to facilitate dispersal.
  4. Add 2.0 to 3.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 4 X 103 to 6 X 103 viable cells/cm2 is recommended.
  7. Incubate cultures at 37.0°C. Subculture when cell density reaches between 5 X 104 and 6 X 104 cells/cm2.
Subcultivation ratio: 1:8 to 1:12 twice weekly Medium renewal: every 2 to 3 days
Cryopreservation Freeze medium: fetal bovine serum (FBS), 90%; DMSO, 10%
Culture Conditions Temperature: 37.0°C Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile CSF1PO: 12 D13S317: 12, 13 D16S539: 12, 13 D5S818: 11 D7S820: 9, 10 TH01: 8, 9 TPOX: 8, 11 vWA: 17 Amelogenin: XY
Population Doubling Level (PDL)

Longevity: >15 PDLs post-cryopreservation recovery
Population Doubling Time approximately 30 hours
Name of Depositor M Ouellette
Year of Origin May 2000
References

Lee KM. et al. Notch2-positive progenitors with the intrinsic ability to give rise to pancreatic ductal cells. Lab. Invest. 85 (8): 1003-1012, 2005. Pubmed: 15924149

Lee KM. et al. Immortalization with telomerase of the Nestin-positive cells of the human pancreas. Biochem. Biophys. Res. Commun. 301(4):1038-1044, 2003. Pubmed: 12589817

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