GMMe [EPI] (ATCC® CRL-2674)

Organism: Mustela vison, mink  /  Cell Type: Epithelial,fibroblast  /  Disease: Endometrial

Organism Mustela vison, mink
Cell Type Epithelial,fibroblast
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain SV40 viral DNA sequences
Disease Endometrial
Age adult
Gender female
Applications
The GMMe (ATCC CRL-2674) and GMMs (ATCC CRL-2675) cell lines were established by stable transfection of endometrial tissue using a plasmid vector, pBAPSV40TtsA58, encoding the SV40 large T antigen driven by the human beta-actin promoter.
The cuboidal GMMe cells exhibit epithelial characteristics and the fibroblast GMMs cells exhibit characteristics consistent with a stromal origin.
The epithelial cells are strongly positive for cytokeratin and weakly positive for vimentin whereas the fibroblast cells are positive for vimentin and negative for cytokeratin.
Both lines are negative for desmin and positive for alkaline phosphatase.
These cell lines are used in coculture with mink embryos in obligate diapause to enhance the length and frequency of embryo survival in vitro.
Storage Conditions liquid nitrogen vapor phase
Derivation
The GMMe (ATCC CRL-2674) and GMMs (ATCC CRL-2675) cell lines were established by stable transfection of endometrial tissue using a plasmid vector, pBAPSV40TtsA58, encoding the SV40 large T antigen driven by the human beta-actin promoter.
Clinical Data
female
Genes Expressed
alkaline phosphatase,cytokeratin,The epithelial cells are strongly positive for cytokeratin and weakly positive for vimentin whereas the fibroblast cells are positive for vimentin and negative for cytokeratin.
Cellular Products
alkaline phosphatase
cytokeratin
Comments
The GMMe (ATCC CRL-2674) and GMMs (ATCC CRL-2675) cell lines were established by stable transfection of endometrial tissue using a plasmid vector, pBAPSV40TtsA58, encoding the SV40 large T antigen driven by the human beta-actin promoter.
The cells were cotransfected with a second plasmid vector, pSV2neo, to confer neomycin resistance. Transfected cells were selected in medium containing G418.
The cuboidal GMMe cells exhibit epithelial characteristics and the fibroblast GMMs cells exhibit characteristics consistent with a stromal origin.
The epithelial cells are strongly positive for cytokeratin and weakly positive for vimentin whereas the fibroblast cells are positive for vimentin and negative for cytokeratin. Both lines are negative for desmin and positive for alkaline phosphatase.
These cell lines are used in coculture with mink embryos in obligate diapause to enhance the length and frequency of embryo survival in vitro.
Complete Growth Medium The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No. 30-2006. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing
Protocol:
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. 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 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 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Name of Depositor BD Murphy
Year of Origin June 13, 1994
References

Moreau GM, et al. Development of immortalized endometrial epithelial and stromal cell lines from the mink (Mustela vison) uterus and their effects on the survival in vitro of mink blastocysts in obligate diapause. Biol. Reprod. 53: 511-518, 1995. PubMed: 7578673

Basic Documentation
References

Moreau GM, et al. Development of immortalized endometrial epithelial and stromal cell lines from the mink (Mustela vison) uterus and their effects on the survival in vitro of mink blastocysts in obligate diapause. Biol. Reprod. 53: 511-518, 1995. PubMed: 7578673