HBE4-E6/E7-C1細胞

器官來源： 肺

數(shù)量： 大量

細胞形態(tài)： 上皮樣

年限： 60 years

細胞類型： 其他細胞類型

是否是腫瘤細胞： 0

物種來源： 人

運輸方式： 凍存運輸

組織來源： bronchus

生長狀態(tài)： 貼壁生長

HBE4-E6/E7-C1細胞ATCC Number： CRL-2079?

Designations: HBE4-E6/E7-C1 [NBE4-E6/E7-C1]

Depositors: J Viallet

Biosafety Level: 2 [Cells contain human papilloma viral DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial

Source: Organ: lung

Tissue: bronchus

Cell Type: HBE4-E6/E7-C1細胞epithelial; human papillomavirus 16 (HPV-16) E6/E7 transformed

Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Cytogenetic Analysis: 45, X, -Y, dup (5), -8, +9, -14, -15, -20, -21, -22, +mar1, +mar2, +der(8q;13q)

Age: 60 years

Gender: male

Ethnicity: Caucasian

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.

Propagation: HBE4-E6/E7-C1細胞ATCC 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: HBE4-E6/E7-C1細胞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.

Preservation: culture medium, 90%; glycerol, 10%

Doubling Time: 24 hrs

References: 22447: 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. HBE4-E6/E7-C1細胞PubMed: 8174640