| BCRJ Code | 0232 |
| Cell Line | T84 |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Colon; Derived From Metastatic Site: Lung |
| Morphology | Epithelial |
| Disease | Coloretal Carcinoma |
| Growth Properties | Adherent |
| Sex | Male |
| Age/Ethinicity | 72 Year / |
| Derivation | The T84 cell line is a transplantable human carcinoma cell line derived from a lung metastasis of a colon carcinoma in a 72-year-old male. Tumor tissue was inoculated subcutaneously and serially transplanted in BALB/c nude mice. |
| Applications | The T84 cell line is a transplantable human carcinoma cell line. It is also a suitable transfection host. |
| DNA Profile | Amelogenin: X CSF1PO: 10 D13S317: 9 D16S539: 10,11 D5S818: 12 D7S820: 8,10 THO1: 6,9 TPOX: 8,11 vWA: 17,18 |
| Tumor Formation: | Yes, in nude mice (Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 10(7) cells) |
| Products | Carcinoembryonic antigen (CEA), 600ng/ml per 10Ecellsper 10 days; Keratin. |
| Biosafety | 1 |
| Addtional Info | The original histological characteristics of the colon carcinoma were maintained throughout transplantation in nude mice. After 23 passages in athymic mice, the T84 cell line was established. These cells grow to confluence as monolayers and exhibit tight junctions and desmosomes between adjacent cells. They have receptors for many peptide hormones and neurotransmitters and maintain vectorial electrolyte transport. This line exhibits tight junctions, and desmosomes between adjacent cells. The cells are positive for keratin by immunoperoxidase staining. |
| Culture Medium | 1:1 mixture of Dulbecco's modified Eagle's medium and F12 Medium containing 1% non-essential amino acids, 2 mM L-glutamine, 1 mM sodium pyruvate and fetal bovine serum to a final concentration of 5%. |
| 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 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 | Twice 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 | Dharmsathaphorn K, et al. A human colonic tumor cell line that maintains vectorial electrolyte transport. Am. J. Physiol. 246: G204-G208, 1984. PubMed: 6141741 Murakami H, Masui H. Hormonal control of human colon carcinoma cell growth in serum-free medium. Proc. Natl. Acad. Sci. USA 77: 3464-3468, 1980. PubMed: 6932031 White LJ, et al. Attachment and entry of recombinant norwalk virus capsids to cultured human and animal cell lines. J. Virol. 70: 6589-6597, 1996. PubMed: 8794293 Cross References: Nucleotide (GenBank) : BE519991 EST-TIG1 cDNA from T84 cells Homo sapiens cDNA, mRNA sequence. |
| Depositors | LILIAN CUPPARI VALLE – UNIFESP-EPM |
| Cellosaurus | CVCL_0555 |
