CHO-1C6
CRL-1793 ™
- Product category
-
Animal cells
- Organism
-
Cricetulus griseus, hamster, Chinese
- Morphology
- epithelial
- Tissue
- Ovary
- Applications
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3D cell culture
- Product format
- Frozen
Documentation
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.
Detailed product information
General
- Specific applications
-
produces protein glucose-6-phosphate isomerase neuroleukin
Characteristics
- Growth properties
- Adherent
- Derivation
-
This cell line was produced by cotransfection with neuroleukin and dihydrofolate reductase transcription units.pAdD26SVp(A)3 was constructed by cloning the mouse dihydrofolate reductase cDNA from the plasmid pAdD26-1 into the vector, pSVOd. A 900 bp BclI/PstI fragment of SV40 containing the early polyadenylation site was inserted.
- Genes expressed
- glucose-6-phosphate isomerase (neuroleukin) [Nlk*]
- Comments
-
Cotransfection and cointegration of pAdNLK and pAdD26SVp(A)3 enables the SV40 enhancer element of pAdNLK to activate the mouse dihydrofolate reductase transcription unit of pAdD26SVp(A)3.This cell line was produced by cotransfection with neuroleukin and dihydrofolate reductase transcription units.Production of neuroleukin was increased by selection with methotrexate.Recently, it has been shown that the polypeptide neuroleukin is in fact glucophosphoisomerase.pAdNLK contains the SV40 enhancer, the mouse neuroleukin cDNA, the mouse dihydrofolate reductase cDNA, the SV40 polyadenylation sequence, and pBR322 sequences minus the inhibitory sequences from 1424 to 2490.pAdD26SVp(A)3 contains the adenovirus major late promoter (AdMLP), the mouse dihydrofolate reductase cDNA, the SV40 polyadenylation sequence, pBR322 sequences minus the inhibitory sequences from 1424 to 2490, and the SV40 origin of replication.The pAdNLK insert contains the complete nucleotide sequence and 5' and 3' untranslated sequences.The cell line CHO-IC6 also contains pAdD26SVp(A)3.Selection of the parental line for growth in methotrexate (MTX) coamplified MTX resistance and neuroleukin secretion. CHO-IC6 secretes 4.4 ug neuroleukin per 10(7) cells per 24 hr.pAdD26SVp(A)3 was constructed by cloning the mouse dihydrofolate reductase cDNA from the plasmid pAdD26-1 into the vector, pSVOd. A 900 bp BclI/PstI fragment of SV40 containing the early polyadenylation site was inserted.
Handling information
- Unpacking and storage instructions
-
- Check all containers for leakage or breakage.
- 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
- Alpha minimum essential medium with 2 mM L-glutamine and 20 nM methotrexate, 90%; fetal bovine serum, 10%
- Handling procedure
-
HANDLING PROCEDURE FOR FROZEN CELLS
- Initiate culture as soon as possible upon receipt.
- Thaw by rapid agitation in 37°C water bath. Thawing should be rapid (within
40-60 seconds). As soon as the ice is melted, remove the ampule from the water
bath and immerse in 70% ethanol at room temperature. All of the operations from
this point on should be carried out under strict aseptic conditions.
- Transfer the cell suspension and dilute it with the recommended culture
medium in a culture flask (see specific batch information above for dilution
ratio); incubate at 37°C with 5% CO2 in air atmosphere. Since it is important
to avoid excessive alkalinity of the medium during recovery of the cells, it is
suggested that the culture medium be placed into the culture flask, tube, etc.
and the pH be adjusted, as necessary, prior to the addition of the ampule
contents. Note that the bicarbonate content of the culture medium will
determine whether an atmosphere containing CO2 will be required.
- It is not necessary to remove the freezing additive. However, if desired, the
culture medium may be changed to remove the protective freezing additive
(dimethylsulfoxide) 24 hours after thawing. If it is desired that the freezing
additive be removed immediately, or that a more concentrated cell suspension be
obtained, centrifuge the above diluted suspension at approximately 125 xg for
10 minutes, discard the fluid and resuspend the cells with growth medium at the
dilution ratio given in the specific batch information above.
- Subculturing procedure
-
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommendedMedium Renewal: 2 to 3 times per weekRemove medium, rinse monolayer with fresh 0.25% trypsin solution, remove trypsin and let the culture sit at room temperature (or 37C) until the cells detach (about 10 minutes). Add fresh medium, aspirate and dispense into new flasks.
Quality control specifications
- Mycoplasma contamination
- Not detected
History
- Depositors
- M Gurney
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.
Frequently Asked Questions
References
Curated Citations
Kaufman RJ, Sharp PA. Construction of a modular dihydrofolate reductase cDNA gene: analysis of signals utilized for efficient expression. Mol. Cell. Biol. 2: 1304-1319, 1982. PubMed: 6131378
Kaufman RJ, et al. Coamplification and coexpression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Mol. Cell. Biol. 5: 1750-1759, 1985. PubMed: 4040603
Gurney ME, et al. Molecular cloning and expression of neuroleukin, a neurotrophic factor for spinal and sensory neurons. Science 234: 566-574, 1986. PubMed: 3764429
Itagaki E, et al. Neuroleukin: a lymphokine product of lectin-stimulated T cells. Science 234: 574-581, 1986. PubMed: 3020690
Chaput M, et al. The neurotrophic factor neuroleukin is 90% homologous with phosphohexose isomerase. Nature 332: 454-455, 1988. PubMed: 3352744
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