MDBK (NBL-1)

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

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.

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.

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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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.

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