NCTC clone 2472

NCTC clone 2472 was derived in May, 1956, by K.K. Sanford, et al. from NCTC strain 1742 (Subline VII) of the parent NCTC clone 1328.

The parent line was derived by G.D. Likely et al. in November, 1950, from a culture of subcutaneous areolar and adipose connective tissue taken from a normal 82-day-old male C3H/HeN mouse.

While cultures of NCTC strain 1742 produced sarcomas in 97% of the mice injected, cultures of NCTC clone 2472 under similar conditions produced sarcomas in 65% of the mice injected.

NCTC clone 2472 has been compared with related clones, such as NCTC clone 2555 (ATCC CCL-12), and certain other cell lines with respect to neoplastic and physiologic properties, nutitional needs, karyology, enzyme activities, and rates of glycolysis.

NCTC clone 2472 cells have been cultured since origin in an antibiotic-free mixture of medium NCTC 109 (135) with 10% horse serum.

Plating efficiency is approximately 10% in the recommended culture medium.

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 75 cm² tissue culture flask and dilute with the recommended complete culture medium (see the specific batch information for the recommended dilution ratio). 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).
4. 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.
5. If it is desired that the cryoprotective agent be removed immediately, or that a more concentrated cell suspension be obtained, centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes. Discard the supernatant and resuspend the cells with fresh growth medium at the dilution ratio recommended in the specific batch information.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Dulbecco's Phosphate Buffered Saline (D-PBS; ATCC 30-2200).
3. Add 2.0 to 3.0 mL of Trypsin-EDTA for Primary Cells (ATCC PCS-999-003) solution to flask and observe cells under an inverted microscope until cell layer is dispersed.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:5 is recommended

Medium Renewal: 3 times per week

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.

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.

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