英文名： Homo sapiens, human│ectocervix; cervix
类型： epithelial HPV-16 E6/E7 transformed
应用领域： These cell lines may provide the basis for valid, reproducible in vitro models for studies on cervicovaginal physiology and infections and for testing pharmacological agents for intravaginal application.
|培养基||Keratinocyte-Serum Free medium (GIBCO-BRL 17005-042) with 0.1 ng/ml human recombinant EGF, 0.05 mg/ml bovine pituitary extract, and additional calcium chloride 44.1 mg/L (final concentration 0.4 mM)|
|传代方法||Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Note: The cells should not be allowed to become confluent, subculture at 60 to 90% of confluence. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.03% (w/v) EDTA solution to remove all traces of serum that contains trypsin inhibitor. 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°C to facilitate dispersal. Neutralize the trypsin by adding 6.0 to 8.0 mL of a 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium containing 10% fetal bovine serum. Aspirate cells by gently pipetting. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh serum-free growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Place culture vessels in incubators at 37°C. Cells will not attach well for 24 hours after subculture. Subcultivation Ratio: 1:4 to 1:6 Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994
|生长条件||Atmosphere: air, 95%; carbon dioxide (CO2), 5%Temperature: 37°C|
|存储条件||liquid nitrogen vapor phase;Freeze medium: A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium, 85%; fetal bovine serum, 10%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase