Lec1 [originally named Pro-5WgaRI3C] (ATCC® CRL-1735)

Organism: Cricetulus griseus, hamster, Chinese  /  Tissue: ovary  / 

Organism Cricetulus griseus, hamster, Chinese
Tissue ovary
Product Format frozen
Morphology epithelial
Culture Properties loosely adherent
Biosafety Level 1
Gender female
Applications This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Derivation
This line is a mutant clone derived from the parental CHO clone Pro-5 (a proline auxotroph, see ATCC CRL-1781) by selection for resistance to wheat germ agglutinin.
Clinical Data
female
Comments
The cells lack GlcNAc glycosyl transferase so that N-linked carbohydrates are blocked at the Man5-GlcNAC2-Asn intermediate. The line may be useful for studying the effects of altered N-linked glycosylation on the function and compartmentalization of endogenous glycoproteins or glycoproteins introduced by viral infection or transfection of foreign DNA.
The population contains Pro+ revertants at low frequency (approximately 1 in 100,000 cells), and should be suitable for complementation studies which require the Pro- marker.
The cells are proline auxotrophs and must be grown in a medium that contains proline (40 mg/L).
The population contains Pro+ revertants at low frequency (approximately 1 in 100000 cells), and should be suitable for complementation studies which require the Pro- marker.
Complete Growth Medium Alpha minimum essential medium with ribonucleosides and deoxyribonucleosides, 90%; fetal bovine serum, 10%
Subculturing
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 mL of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Name of Depositor P Stanley
References

Stanley P, et al. Selection and characterization of eight phenotypically distinct lines of lectin-resistant Chinese hamster ovary cells. Cell 6: 121-128, 1975. PubMed: 1182798

Stanley P, et al. Chinese hamster ovary cells selected for resistance to the cytotoxicity of phytohemagglutinin are deficient in a UDP-N-acetylglucosamine-- glycoprotein N-acetylglucosaminyltransferase activity. Proc. Natl. Acad. Sci. USA 72: 3323-3327, 1975. PubMed: 1059116

Cross References

Nucleotide (GenBank) : U65792 Cricetulus griseus mutant N-acetylglucosaminyltransferase I mRNA, complete cds.

Basic Documentation
References

Stanley P, et al. Selection and characterization of eight phenotypically distinct lines of lectin-resistant Chinese hamster ovary cells. Cell 6: 121-128, 1975. PubMed: 1182798

Stanley P, et al. Chinese hamster ovary cells selected for resistance to the cytotoxicity of phytohemagglutinin are deficient in a UDP-N-acetylglucosamine-- glycoprotein N-acetylglucosaminyltransferase activity. Proc. Natl. Acad. Sci. USA 72: 3323-3327, 1975. PubMed: 1059116