| BCRJ Code | 0365 |
| Cell Line | KLE |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Uterus/endometrium |
| Morphology | Epithelial |
| Disease | Adenocarcinoma |
| Growth Properties | Adherent |
| Sex | Female |
| Age/Ethinicity | 64 Year / Caucasian |
| Tumor Formation: | Yes, Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 107 cells. |
| Biosafety | 1 |
| Addtional Info | Electron microscopy of tumors formed in nude mice shows microvilli and junctional complexes, and nucleolar channel systems are present that are similar to those seen in normal endometrium under progestational stimulation. The tumors do not form glands. |
| Culture Medium | DMEM: F-12 Medium contains 2.5 mM L-glutamine, 0.5 mM sodium pyruvate with fetal bovine serum to a final concentration of 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. Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until the 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. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Place culture vessels in incubators at 37°C. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010. |
| Subculturing Medium Renewal | Twice per week |
| Subculturing Subcultivation Ratio | 1:2 to 1:3 is recommended |
| Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
| Cryopreservation | 95% FBS + 5% DMSO (Dimethyl sulfoxide) |
| Thawing Frozen Cells | SAFETY PRECAUTION:
It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6). |
| References | Hendricks DT, et al. FHIT gene expression in human ovarian, endometrial, and cervical cancer cell lines. Cancer Res. 57: 2112-2115, 1997. PubMed: 9187105 |
| Depositors | Etel Rodrigues Pereira Gimba - Instituto Nacional de Câncer |
| Cellosaurus | CVCL_1329 |
