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ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
Cells had contained Bovine viral diarrhea virus (BVDV)
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may 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 vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
The cells were originally obtained in July, 1967 at passage 96 from the ATCC as a frozen amplule (F-145). The cells were found to be contaminated with BVD and could not be used in the production of CCL-22.
The cell line was submitted to the American Type Culture Collection by R.A. Van Deusen in passage 110 in August, 1982. This deposit was tested found to be BVD free. The cells deposited in 1982 were the ones used in the production of the CCL-22 cell line.
A specific lot of CCL-22, MDBK (NBL-1) bovine kidney cells was found to be positive for bovine viral diarrhea virus (BVDV), following an investigation by ATCC (Dec. 2015). Testing for BVDV was performed in compliance with the Code of Federal Regulations, Title 9, Section 113.52 (E &F) using virus-specific fluorescent antibody (FA) technique as well as non-specific tests for hemadsorption and cytopathic effects (CPE). Samples were found to be positive for BVDV by FA. Hemadsorption and CPE were not observed in the sample inoculated cultures. As a result, the BSL status for CCL-22 was changed to “2”.
Subsequent lots of CCL-22, MDBK (NBL-1) bovine kidney cells are tested for BVDV and the results will be indicated on the Certificate of Analysis for the lot.
The cells are positive for keratin by immunofluorescence
To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
Rinse the cell sheet twice with fresh 0.25% trypsin, 0.03% EDTA solution, and remove the trypsin solution. Allow the flask to stand at room temperature for 10 to 15 minutes, add fresh medium, aspirate and dispense into new flasks.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
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Madin SH, Darby NB Jr.. Established kidney cell lines of normal adult bovine and ovine origin. Proc. Soc. Exp. Biol. Med. 98: 574-576, 1958. PubMed: 13567776
Bolin SR, et al. Survey of cell lines in the American Type Culture Collection for bovine viral diarrhea virus. J. Virol. Methods 48: 211-221, 1994. PubMed: 7989438
Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321
Russell DW, Miller AD. Foamy virus vectors. J. Virol. 70: 217-222, 1996. PubMed: 8523528
USEPA Manual of Methods for Virology - EPA publication. Washington, DC:Environmental Protection Agency;EPA EPA 600/4-84/013 (R9), 2001