CHO-1C6 (ATCC® CRL-1793)

Organism: Cricetulus griseus, hamster, Chinese  / 

Organism Cricetulus griseus, hamster, Chinese
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Applications
produces protein glucose-6-phosphate isomerase neuroleukin
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*],glucose-6-phosphate isomerase
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.
Complete Growth Medium Alpha minimum essential medium with 2 mM L-glutamine and 20 nM methotrexate, 90%; fetal bovine serum, 10%
Subculturing
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week
Remove 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.
Name of Depositor M Gurney
References

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

Mark Gurney, personal communication

Basic Documentation
References

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

Mark Gurney, personal communication