| BCRJ Code | 0339 |
| Cell Line | HOS |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Bone |
| Morphology | Mixed, fibroblast and epithelial like cells |
| Disease | Osteosarcoma |
| Growth Properties | Adherent |
| Sex | Female |
| Age/Ethinicity | 13 Year / Caucasian |
| Biosafety | 1 |
| Addtional Info | HOS cells exhibit flat morphology, low saturation density, low plating efficiency in soft agar and are sensitive to chemical and viral transformation. |
| Culture Medium | Dulbecco's Modified Eagle's Medium (DMEM) with 1% non-essential amino acids, 2 mM L-glutamine and 10% of fetal bovine serum. |
| 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 | 2 to 3 times per week |
| Subculturing Subcultivation Ratio | 1:2 to 1:4 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 | Rhim JS, et al. Non-producer human cells induced by murine sarcoma virus. Int. J. Cancer 15: 23-29, 1975. PubMed: 165148 McAllister RM, et al. Cultivation in vitro of cells derived from a human osteosarcoma. Cancer 27: 397-402, 1971. PubMed: 5100401 Rhim JS, et al. Characterization of non-producer human cells induced by Kirsten sarcoma virus. Int. J. Cancer 16: 840-849, 1975. PubMed: 171229 Rhim JS. Characterization of sarcoma-positive, leukemia-negative (S+L-) human cells induced by the feline leukemia virus pseudotype of Moloney sarcoma virus. Proc. Soc. Exp. Biol. Med. 167: 597-606, 1981. PubMed: 6269117 Rhim JS, et al. Differential susceptibility of human cells to transformation by murine and avian sarcoma viruses. Proc. Soc. Exp. Biol. Med. 170: 350-358, 1982. PubMed: 6283561 . . Nat. New Biol. 230: 279-282, 1971. Yee A, et al. Biochemical characterization of the human cyclin-dependent protein kinase activating kinase. J. Biol. Chem. 271: 471-477, 1996. PubMed: 8550604 |
| Depositors | Banco de Células do Rio de Janeiro |
| Cellosaurus | CVCL_0312 |
