ATCC ATCC Logo 0
  • Quick Order

C6/lacZ7

CRL-2303

Product category
Animal cells
Product type
Reporter-labeled cell
Organism
Rattus norvegicus, rat
Cell type
glial cell
Morphology
fibroblast
Tissue
Brain
Disease
Glioma
Applications
3D cell culture
Assay development
High-throughput screening
Neuroscience
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
Buy Now
Price: $791.00 EA
Discounts may be available for our fellow nonprofit organizations.

Generally ships within 1-3 business days

Documentation

Biosafety Icon Biosafety Level 1

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.

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
C6/lacZ7 is a subclone of the C6/lacZ cell line (ATCC CRL-2199) that was established in 1989 from the N-nitrosomethylurea-induced glial rat, outbred strain, C6 cell line.
The cells were cultured in G418 for 14 days, cloned, and evaluated for beta-gal production.
The cells constitutively express the lacZ reporter gene product, E. coli derived beta-gal, as revealed on tissue sections by histochemical stain, and single tumor cells can be identified.
Lymphocytes and other responding cells can be identified by double labeling with antibodies on the same slide.
The contrast between stained cells and background facilitates image analysis.
The depositor has confirmed that the C6/lacZ7 subclone grows in rat brain, and that the lacZ marker can be detected by histochemistry for b-gal.
This is one of few models that permit quantitative analysis of microscopic tumor in the brain.
The beta-gal expression is less stable than for 9L/lacZ. Cells should be used soon after thawing or re-cloned.
Methods for the use of C6/lacZ may be found in Cancer Res.

Characteristics

Growth properties
Adherent
Derivation
C6/lacZ7 is a subclone of the C6/lacZ cell line (ATCC CRL-2199) that was established in 1989 from the N-nitrosomethylurea-induced glial rat, outbred strain, C6 cell line.
The cells constitutively express the lacZ reporter gene product, E. coli derived beta-gal, as revealed on tissue sections by histochemical stain, and single tumor cells can be identified.
Strain
outbred
Tumorigenic
Yes;
Yes, in the brains of CD Fischer 344 rats
Genes expressed
beta galactosidase (beta-gal)
Comments
C6/lacZ7 is a subclone of the C6/lacZ cell line (ATCC CRL-2199) that was established in 1989 from the N-nitrosomethylurea-induced glial rat, outbred strain, C6 cell line.
C6 cells were infected with the BAG replication deficient retroviral vector carrying the E. coli lacZ gene encoding beta-gal and the Tn5 neomycin gene, which confers resistance to G418.
The cells were cultured in G418 for 14 days, cloned, and evaluated for beta-gal production.
To obtain the subclone, C6/lacZ was cloned in 1996 at limiting dilution.
The cells constitutively express the lacZ reporter gene product, E. coli derived beta-gal, as revealed on tissue sections by histochemical stain, and single tumor cells can be identified.
Lymphocytes and other responding cells can be identified by double labeling with antibodies on the same slide.
The contrast between stained cells and background facilitates image analysis.
The subclone was chosen because it has a more stable expression of the lacZ marker. The depositor has confirmed that the C6/lacZ7 subclone grows in rat brain, and that the lacZ marker can be detected by histochemistry for b-gal.
This is one of few models that permit quantitative analysis of microscopic tumor in the brain.
The tumor mimics important features of human brain tumor growth and spread.
The beta-gal expression is less stable than for 9L/lacZ. Cells should be used soon after thawing or re-cloned.
The cloned cells have been shown to retain the marker for up to eleven weeks in culture and up to three weeks in the brain.
Methods for the use of C6/lacZ may be found in Cancer Res. 52: 1018-1025, 1992; ibid., 53: 176-182, 1993.

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 Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • O.1 mM Non-Essential Amino Acids (NEAA)
  • fetal bovine serum to a final concentration of 10%
Temperature
37°C
Atmosphere
95% Air, 5% CO2
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 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.

Note: 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.

Subculturing procedure
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  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: 1:6 to 1:12
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

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

Quality control specifications

Mycoplasma contamination
Not detected

History

Depositors
LA Lampson

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.

Permits & Restrictions

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

Lampson LA, et al. Exploiting the lacZ reporter gene for quantitative analysis of disseminated tumor growth within the brain: use of the lacZ gene product as a tumor antigen, for evaluation of antigenic modulation, and to facilitate image analysis of tumor growth in situ. Cancer Res. 53: 176-182, 1993. PubMed: 8416743

Lampson LA, et al. Disseminating tumor cells and their interactions with leukocytes visualized in the brain. Cancer Res. 52: 1018-1025, 1992. PubMed: 1737331

Lampson LA, et al. A new model permits study of the immune response to individual tumor cells within the brain. Neurology 40: 396-397, 1990.

Dutta T, et al. Robust ability of IFN-gamma to upregulate class II MHC antigen expression in tumor bearing rat brains. J. Neuro-Oncol. 64: 31-44, 2003. PubMed: 12952284

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

View All Curated Citations for this Product

Frequently Asked Questions

Need assistance with this product? Contact our Technical Support team.

Telephone

Telephone

US and Puerto Rico

800-638-6597

Outside the US

+1-703-365-2700
hours

Hours of Operation

Monday-Friday
9:30am - 5:30pm
US Eastern Time