| BCRJ Code | 0412 |
| Cell Line | RPMI 2650 |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Nose; Nasal septum |
| Cell Type | Epithelial |
| Morphology | Epithelial |
| Disease | Squamous Cell Carcinoma |
| Growth Properties | Adherent |
| Sex | Male |
| Age/Ethinicity | 52 Year / |
| Derivation | From the pleural effusion of an extensive malignant tumour of the nasal septum, diagnosed as anaplastic squamous cell carcinoma. Nearly normal karyotype, stability of chromosome number and probable origin from malignant cells make the cell line unique amongst permanent human cell lines. Cells are very small and grow in clusters fusing to form thick layers. |
| Virus Succeptility: | Human poliovirus 1 Herpes simplex virus Vesicular stomatitis, Glasgow (Indiana) Vesicular stomatitis, Orsay (Indiana) |
| Products | Genes expressed: mucoid; keratin Isoenzymes: G6PD, B |
| Biosafety | 1 |
| Culture Medium | EMEM (EBSS) + 2mM Glutamine + 1% Non Essential Amino Acids (NEAA) + 10% of fetal Bovine Serum. |
| Subculturing | This cells attach in clusters. Cells will pile and the culture does not get 100% confluent. Volumes used in this protocol are for 75 cm2 flasks; 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 which contains trypsin inhibitor. 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. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures 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 | 2 to 3 times per week |
| Subculturing Subcultivation Ratio | 1:2 to 1:4 |
| 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 | Exp Cell Res 1965;39:190; Cancer 1964;17:170 |
| Depositors | Banco de Células do Rio de Janeiro |
| Cellosaurus | CVCL_1664 |
