| BCRJ Code | 0095 |
| Cell Line | GT1-7 |
| Species | Mus musculus |
| Vulgar Name | Mouse |
| Tissue | Brain |
| Cell Type | Fibroblast |
| Morphology | Fibroblast |
| Growth Properties | Adherent |
| Derivation | The GT1-7 line was immortalized by expression of SV40 T antigen oncogene in hypothalamic GnRH neurons. |
| Products | GnRH - Gonadotropin-Releasing Hormone. |
| Biosafety | 2 |
| Addtional Info | GT1-7 cell line is sensitive to trypsin and low confluence. |
| Culture Medium | Dulbecco's modified Eagle's medium with 4.5 g/L glucose and 10% of fetal bovine serum. |
| Subculturing | 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 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. Doubling time: 36 hours 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 | Every 2 to 3 days |
| Subculturing Subcultivation Ratio | 1:5 to 1:7 |
| 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 | Nelson SB, LAwson MA, Kelley CG, Mellon PL (2000) "neuron-specific expression of the rat gonadotropin-releasing hormone gene is confered by interactions of a defined promoter with the enhacer in GT1-7 cells". Mol Endocrinol 14 (9): 1509-22. Mellon, P.L., Windle, J,J., Goldsmith, P., Pedula, C., Robets, J. and Weiner, R.I. 1990. Imortalization of hypothalamic GnRH neurons by genetically targeted tumorigenesis. Neuron 5: 1-10. Liposits, Z., Mechenthaler,I., Westel, W.C., Reid, J. J., Mellon, P. L., Weiner, R.I. and Negro-Villar, A.1991. Morphological chaacterization of immortalized hypothalamic neurons synthesizing luteinizing hormone-releasing hormone. Endocrinol. 129: 1575-1583. Wetsel, W.C., Valença, M.M., Merchenthaler, I. Liposits, Z., López, F.J., Weiner, R.I., Mellon, P.L. and Negro-Villar, A. 1992. Intrinsic pulsatile secretory activity of immortalized luteinizing hormone-releasing hormone-secreting neurons. Proc. Natl. Acad. Sci. USA 89:4149-4153. Whyte, D. B., Lawson, M.A., Belsham,D. D., Eraly, S.A. Bond, C.T., Adelman, J. P., and Mellon, P.L. 1995. A Neuron-Specific Enhancer Targets Expression of the Gonadotropin-Releasing Hormone Gene to Hypothalamic Neurosecretory Neurons. Molecular Endocrinology 9, 467-477. |
| Depositors | JOSE BARRETO CAMPELLO CARVALHEIRA; UNICAMP |
| Cellosaurus | CVCL_0281 |
