||Aedes albopictus clone C6/36
||Mosquito, Asian Tiger
||This cell line is useful for the replication of flaviviruses and reportedly can be used to replicate Dengue viruses to high titers. They are also a suitable transfection host
||Yes, did form colonies in semisolid medium.
No, the cells were not tumorigenic in immunosuppressed mice.
||The cells are non-anchorage dependent, are not tumorigenic and maintain a diploid chromosome number.
||Dulbecco's Modified Eagle's Medium (DMEM) modified to contain 2 mM L-glutamine, 4500 mg/L glucose and fetal bovine serum to a final concentration of 10%.
||Subcultures are prepared by scraping or by vigorous pipetting.
Remove the old medium, add fresh complete culture medium, dislodge cells from the floor of the flask, aspirate and dispense into new flasks.
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
||Twice per week
|Subculturing Subcultivation Ratio
||1:4 to 1:10 is recommended
||Atmosphere: air, 95%; carbon dioxide (CO2), 5%
||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).
||Igarashi A. Isolation of a Singh's Aedes albopictus cell clone sensitive to Dengue and Chikungunya viruses. J. Gen. Virol. 40: 531-544, 1978. PubMed: 690610
Singh KRP. Cell cultures derived from larvae of Aedes albopictus (Skuse) and Aedes aegypti (L.). Curr. Sci. 36: 506-508, 1967.
Clone C6/36 was derived from A. albopictus cells (see ATCC CCL-126) which were adapted to Eagle's minimum essential medium by A. Igarashi and then cloned and recloned by seeding single cell suspensions into petri dishes.
||Danielle da Glória de Souza - UFMG