Lec8 [originally named Pro-5WgaRVIII3D] (ATCC® CRL-1737)

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

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Organism Cricetulus griseus, hamster, Chinese
Tissue ovary
Product Format frozen
Morphology epithelial
Culture Properties monolayer
Biosafety Level 1
Gender female
Applications

May be useful for studying the role of galactose residues in the function and compartmentalization of glycosylated macromolecules due to a drastic reduction in the transport of UDP-galactose into the Golgi compartment.

Can be used as a Pro- marker in complementation studies due to Pro+ occuring at a high frequency in the population.
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.
Comments The cells exhibit a drastic reduction in the transport of UDP-galactose into the Golgi compartment.

The cells are proline auxotrophs and must be grown in a medium that contains proline (40 mg/L).

The population contains Pro+ revertants at high frequency (approximately 1 in 250 cells).

These cells may also be grown as a suspension culture if agitated. Keep the cell concentration between 2 x 104 and 1 x 106 cells/mL.
Complete Growth Medium Alpha minimum essential medium, 90%; fetal bovine serum, 10%
Subculturing 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. 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 10 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.
  3. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  4. Add appropriate aliquots of the cell suspension to new culture vessels. 
  5. Incubate cultures at 37°C.
ALTERNATIVELY, cultures also can be grown as a suspension and maintained by addition or replacement of fresh medium. Start cultures at 2 x 104 cells/mL and maintain between 2 x 104 and 1 x 106 cells/mL.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: 2 to 3 times per week
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.  
Culture Conditions
Temperature: 37°C
Name of Depositor P Stanley
References

Stanley P. Selection of specific wheat germ agglutinin-resistant (WgaR) phenotypes from Chinese hamster ovary cell populations containing numerous lecR genotypes. Mol. Cell. Biol. 1: 687-696, 1981. PubMed: 9279382

Deutscher SL, Hirschberg CB. Mechanism of galactosylation in the Golgi apparatus. A Chinese hamster ovary cell mutant deficient in translocation of UDP-galactose across Golgi vesicle membranes. J. Biol. Chem. 261: 96-100, 1986. PubMed: 3510203

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Stanley P. Selection of specific wheat germ agglutinin-resistant (WgaR) phenotypes from Chinese hamster ovary cell populations containing numerous lecR genotypes. Mol. Cell. Biol. 1: 687-696, 1981. PubMed: 9279382

Deutscher SL, Hirschberg CB. Mechanism of galactosylation in the Golgi apparatus. A Chinese hamster ovary cell mutant deficient in translocation of UDP-galactose across Golgi vesicle membranes. J. Biol. Chem. 261: 96-100, 1986. PubMed: 3510203