0245 - Vero

BCRJ Code 0245
Cell Line Vero
Species Chlorocebus sabaeus
Vulgar Name Monkey; African Green Monkey
Tissue Kidney
Morphology Epithelial
Disease Normal
Growth Properties Adherent
Age/Ethinicity ADULT /
Derivation The Vero cell line was established from kidney of normal adult African green monkey at the Chiba University, Japan. It was brought to NIH by Dr Simizu. This ce4ll line was extensively used in virus replication studies and plaque assays. It has been
Applications This cell line can be used for the detection of verotoxin. This cell line can be used for efficacy testing. This cell line can be used to study malaria biology. This cell line can be used for media testing. This cell line can be used for mycoplasma testing. This cell line is a suitable transfection host. This cell line can be used for the detection of virus in ground beef.
Virus Succeptility: Human poliovirus 1 Human poliovirus 2 Human poliovirus 3 Getah virus Pixuna virus Ross River virus Semliki Forest virus Kokobera virus Modoc virus Guaroa virus Tacaribe virus , Tacaribe virus SV-5 (parainfluenza type 2) SV40 virus Measles virus Rubella virus , Rubella virus Reovirus type 2 Reovirus 3 Simian adenovirus 3 Simian adenovirus 17 Simian adenovirus 11 Simian adenovirus 1 Simian adenovirus 20 Simian adenovirus 20 Simian adenovirus 18 Simian adenovirus 16 Simian adenovirus 8 Simian adenovirus 17 Simian adenovirus 19 Simian adenovirus 21 Simian adenovirus 25 Simian adenovirus 22 Simian adenovirus 23 Simian adenovirus 38 Simian adenovirus 37 Simian adenovirus 27 Simian adenovirus 39 Simian adenovirus 32 Simian adenovirus 34 Simian adenovirus 31 Simian adenovirus 33 Simian adenovirus 36
Virus Resistance: Apeu; Ossa
Biosafety 1
Culture Medium Dulbecco's Modified Eagle's Medium (DMEM) with 2 mM L-glutamine, 1.0 g/L glucose and 10% of fetal bovine serum.
Subculturing Volumes used in this protocol are for 75 cm2 flask; 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 PBS without calcium and magnesium 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. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010.
Subculturing Medium Renewal Every 2 to 3 days
Subculturing Subcultivation Ratio 1:3 to 1:6
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes). 2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. 3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
References Nippon Rinsho 21: 1209-, 1963; Proc. Soc. Exp. Biol. Med. 125: 119-, 1967.
Depositors Banco de Células do Rio de Janeiro