HuZP3-CHOLec3.2.8.1 (ATCC® CRL-2866)

Organism: Cricetulus griseus, hamster, Chinese  / 

Permits and Restrictions

View Permits

Organism Cricetulus griseus, hamster, Chinese
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells containing SV40 viral DNA sequences
Gender female
Applications
HuZP3-CHOLec3.2.8.1 cells express recombinant human ZP3. The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization and in the mouse is composed of three glycoproteins (ZP1, ZP2, ZP3).
Storage Conditions liquid nitrogen vapor phase
Images
Clinical Data
female
Genes Expressed
human ZP3 (zona pellucida glycoprotein 3)
Comments
HuZP3-CHOLec3.2.8.1 cells express recombinant human ZP3. The zona pellucida is an extracellular matrix that mediates taxon-specific fertilization and in the mouse is composed of three glycoproteins (ZP1, ZP2, ZP3). Full length human ZP3 cDNA was cloned into expression vector pEF1/V5-His and transfected into glycosylation-deficient CHO cells. It was compared to mouse ZP3 and found to be quite similar [PubMed: 15379548].
Complete Growth Medium Alpha minimum essential medium without ribonucleosides and deoxyribonucleosides, 90%; fetal bovine serum, 10%
Subculturing
Protocol: Volumes used in this protocol are for 75 sq cm flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 1.0 to 2.0 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 37C to facilitate dispersal.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 3 X 10(3) to 5 X 10(3) viable cells/sq. cm is recommended.
  7. Incubate cultures at 37C. We recommend that you maintain cultures at a cell concentration between 8 X 10(4) and 1.5 X 10(5) cells/sq. cm.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: Complete Growth Medium, 95%; DMSO,5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: 5% CO2 in air recommended
Temperature: 37.0°C
Population Doubling Time about 10 hours
Name of Depositor J Dean
References

Zhao M, et al. Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. Biochemistry 43: 12090-12104, 2004. PubMed: 15379548

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
Other Documentation
References

Zhao M, et al. Mass spectrometry analysis of recombinant human ZP3 expressed in glycosylation-deficient CHO cells. Biochemistry 43: 12090-12104, 2004. PubMed: 15379548