CCL-1 ™
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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.
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.
This cell line has been tested and found negative for ectromelia virus (mousepox).
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.
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 If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.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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Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
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.
Cells that are properly frozen using an effective cryoprotective agent can be stored in liquid nitrogen indefinitely without affecting recovery. Cells of NCTC clone 929 [L cell, L-929, derivative of Strain L] (ATCC CCL-1) were cryopreserved in February 1962. After decades in liquid nitrogen or its vapor, their viability has not significantly declined.
Cell lines maintained on dry ice or in -70°C mechanical freezers, however, often lose their viability very quickly. The loss of viability will, of course, vary from cell line to cell line. Studies done at ATCC show some mouse lines dropping to 0% viability within six months on dry ice and some human lines were non-viable in only four months. Thus, it is recommended that ampoules and vials be thawed as soon as possible upon receipt. If the cells cannot be thawed and cultured immediately, the ampoules and vials should be stored at temperatures below -135°C, preferably in liquid nitrogen vapor.
Morphology can be influenced by the medium used to culture the cells. Variations in growth factors and the presence or absence of attachment factors can affect how cells appear on the substrate. For example, NCTC clone 929 [L cell, L-929, derivative of Strain L] (ATCC CCL-1) is a fibroblast line that appears more epithelial-like under ATCC standard conditions of growth.
Not all cell lines can be adapted to suspension growth. In general, normal diploid, anchorage-dependent (must be attached to a substrate to grow) cells will grow in suspension only with the use of microcarrier beads. However, some cell lines such as L-929 (ATCC CCL-1), HeLa (ATCC CCL-2) and BHK-21 (ATCC CCL-10), which are not anchorage-dependent, can be adapted and variants that grow in suspension already exist.
Patience, and the use of specially modified suspension culture medium, are the keys to adapting cells to suspension growth. Cultures are usually grown in small (250 ml to 1,000 ml) spinner cultures (with half that volume of actual medium), in glass vessels with a stirring paddle suspended inside that is driven at approximately 50 to 100 rpm by a magnetic stirrer. The spinner/suspension culture medium, such as Joklik's modified MEM medium, usually omits calcium and magnesium ions to help prevent clumping and may require an antifoaming agent to prevent foaming if serum is used. The glass culture vessel is usually coated with a siliconizing compound to prevent the cells from sticking to the glass.
Initially, most of the cells will plate out on the glass surface of the vessel or form large clumps with each other. At each passage, those cells that are in suspension are used to inoculate the next vessel. With time, a population of cells may be selected that does not self-aggregate or adhere to a growth surface as readily as the parental line. However, the newly selected line may have lost or acquired characteristics that are different from the original cell population. In most cases, it will be necessary to maintain the culture in suspension with mechanical stirring.
The procedure below was developed for BHK-21 cells, but can be used as a starting point for most cell lines.
For further detail on suspension cell culture techniques, please consult the following references.
Kazazian HH Jr., et al. Restriction site polymorphism in the phosphoglycerate kinase gene on the X chromosome. Hum. Genet. 66: 217-219, 1984. PubMed: 6325324
Fisher EM, et al. Homologous ribosomal protein genes on the human X and Y chromosomes: escape from X inactivation and possible implications for Turner syndrome. Cell 63: 1205-1218, 1990. PubMed: 2124517
Sanford KK, et al. The growth in vitro of single isolated tissue cells. J. Natl. Cancer Inst. 9: 229-246, 1948.
Sugarman BJ, et al. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science 230: 943-945, 1985. PubMed: 3933111
ASTM International Standard Practice for Direct Contact Cell Culture Evaluation of Materials for Medical Devices. West Conshohocken, PA:ASTM International;ASTM Standard Test Method F 0813-07.
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