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MDBK (NBL-1)

CCL-22

Product category
Animal cells
Organism
Bos taurus, cow
Morphology
epithelial
Tissue
kidney
Applications
3D cell culture
High-throughput screening
Toxicology
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

Biosafety Icon BSL 2

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.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific applications
This cell line is suitable as a transfection host

Characteristics

Growth properties
Adherent
Passage history

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.   

Derivation
The MDBK cell line was derived from a kidney of an apparently normal adult steer, February 18, 1957, by S.H. Madin and N.B. Darby. The cells were originally obtained in July, 1967 at passage 96 from the ATCC as a frozen ampule (F-145).­
Age
adult
Gender
Male
Karyotype
Chromosome Frequency Distribution 50 Cells: 2n = 60. The stemline chromosome number is hypodiploid with 2S component occurring at 5%., There were a total of 11-14 marker chromosomes (4-5 metacentric, 3-4 submetacentric and 4-5 acro-telocentric) common to most hypodiploid metaphases. The X was monosomic. Neither HSR chromosomes nor DM's were seen.
Virus susceptibility
Vesicular stomatitis, Orsay (Indiana)
Infectious bovine rhinotracheitis
Bovine parvovirus
Bovine adenovirus 2
Bovine adenovirus 3
Bovine viral diarrhea virus 1
Parainfluenza 3
Genes expressed
keratin
Comments

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

Technical information
ATCC Technical Services does not have technical information on patent deposits that are not produced or characterized by ATCC. Additional information can be found in the corresponding patent available from the patent holder or with the U.S. and/or international patent office.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: horse serum to a final concentration of 10%.
Temperature
37°C
Handling procedure

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.

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring 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. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete growth medium and spin at approximately 125 x g for 5 to 7 minutes.
  4. Resuspend cell pellet with the recommended complete growth medium (see the specific batch information for the culture recommended dilution ratio). and dispense into a 25 cm2 or a 75 cm2 culture flask.  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 complete 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).
  5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.
Subculturing procedure

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

Medium Renewal: Twice per week

Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected

History

Deposited as
Bos taurus
Depositors
S Madin, NB Darby
Patent depository
This material was deposited with the ATCC Patent Depository to fulfill U.S. or international patent requirements. This material may not have been produced or characterized by ATCC.  As an International Depository Authority (IDA) for patent deposits, ATCC is required to complete viability testing only at time of initial deposit of patent material. Patent deposits are made available on behalf of the Depositor when the pertinent U.S. or international patent is issued, but material may not be used to infringe the patent claims.
Year of origin
1957
Cross references
GenBank AF025996 Bos taurus supervillin mRNA, complete cds.
GenBank U64997 Bos taurus ribonuclease K6 gene, partial cds.
GenBank NC_002531 Alcelaphine herpesvirus 1, complete genome.
GenBank AJ238405 Bos taurus mRNA for elongation factor 1 alpha.
GenBank J05217 Bovine beta-1->4-galactosyltransferase mRNA, 5' end.
GenBank AF005370 Alcelaphine herpesvirus 1 L-DNA, complete sequence.

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

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.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Disclosures
This material is cited in a US and/or international patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Depositor of the party to which the material was furnished.

Permits & Restrictions

United States Veterinary Permit for Importation and Transportation of Controlled Materials and Organisms and Vectors

For every order of this item, you must provide a valid Permit for Importation and Transportation of Controlled Materials and Organisms and Vectors (VS Form 16-6A) obtained from the United States Department of Agriculture (USDA), Animal and Plant Health Inspection Service. We cannot ship this item until we receive this permit.

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

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

Frequently Asked Questions

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