In Vitro Technologies :: Products

Permits and Restrictions	View Permits
Organism	Homo sapiens, human
Cell Type	epithelial; human papillomavirus 16 (HPV-16) E6/E7 transforme
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	2 Cells contain human papilloma 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	60 years
Gender	male
Ethnicity	Caucasian

Karyotype	45, X, -Y, dup (5), -8, +9, -14, -15, -20, -21, -22, +mar1, +mar2, +der(8q;13q)
Derivation	The HBE4-E6/E7-C1 (formerly NBE4-E6/E7-C1) cell line was cloned from passage 22 of HBE4-E6/E7 (see ATCC CRL-2078) by limiting dilution.
Clinical Data	male Caucasian 60 years
Comments	The HBE4-E6/E7-C1 (formerly NBE4-E6/E7-C1) cell line was cloned from passage 22 of HBE4-E6/E7 (see ATCC CRL-2078) by limiting dilution. The cloned line is more sensitive than the parental line to induction of terminal differentiation by phorbol esters. The cells are non-viable in DMSO and should be frozen in culture medium with 10% glycerol.

Complete Growth Medium	The base medium for this cell line is provided by Invitrogen (GIBCO) as part of a kit: Keratinocyte Serum Free Medium (K-SFM), Kit Catalog Number 17005-042. This kit is supplied with two additives required to grow this cell line (bovine pituitary extract (BPE) and human recombinant epidermal growth factor (EGF). To make the complete growth medium, you will need to add the following components to the base medium: 0.05 mg/ml BPE - provided with the K-SFM kit 5 ng/ml EGF - provided with the K-SFM kit 10 ng/ml cholera toxin - not provided with kit NOTE: Do not filter complete medium
Subculturing	Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: Twice per week Protocol: Volumes used in this protocol are for 75 sq cm flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution containing 0.5% polyvinylpyrrolidone (PVP). Add 2.0 to 3.0 ml of Trypsin-EDTA-PVP 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 37C to facilitate dispersal. Add 2.0 to 3.0 ml of complete growth medium containing 0.1% soybean trypsin inhibitor and 0.1% bovine serum albumin and aspirate cells by gently pipetting.Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. Subculture before the cells become confluent.
Cryopreservation	culture medium, 90%; glycerol, 10%

Population Doubling Time	24 hrs

Name of Depositor	J Viallet
Passage History	The HBE4-E6/E7-C1 (formerly NBE4-E6/E7-C1) cell line was cloned from passage 22 of HBE4-E6/E7 (see ATCC CRL-2078) by limiting dilution.
References	Viallet J, et al. Characterization of human bronchial epithelial cells immortalized by the E6 and E7 genes of human papillomavirus type 16. Exp. Cell Res. 212: 36-41, 1994. PubMed: 8174640

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